Nucleic acid molecules and other molecules associated with the sucrose pathway

ABSTRACT

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean plants associated with the sucrose pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Nos.: 60/067,000 filed Nov. 24, 1997; 60/069,472 filed Dec. 9, 1997; 60/072,888 filed Jan. 27, 1998; 60/074,201 filed Feb. 10, 1998; 60/074,282 filed Feb. 10, 1998; 60/074,280 filed Feb. 10, 1998; 60/074,281 filed Feb. 10, 1998; 60/074,566 filed Feb. 12, 1998; 60/074,567 filed Feb. 12, 1998; 60/074,565 filed Feb. 12, 1998; 60/075,462 filed Feb. 19, 1998; 60/074,789 filed Feb. 19, 1998; 60/075,459 Feb. 19, 1998; 60/075,461 filed Feb. 19, 1998; 60/075,464 filed Feb. 19, 1998; 60/075,460 filed Feb. 19, 1998; 60/075,463 Feb. 19, 1998; 60/076,912 filed Mar. 6, 1998; 60/077,231 filed Mar. 9, 1998; 60/077,229 filed Mar. 9, 1998; 60/077,230 filed Mar. 9, 1998; 60/078,368 filed Mar. 18, 1998; 60/080,844 filed Apr. 7, 1998; 60/083,067 filed Apr. 27, 1998; 60/083,386 filed Apr. 29, 1998; 60/083,387 Apr. 29, 1998; 60/083,388 filed Apr. 29, 1998; 60/083,389 filed Apr. 29, 1998; 60/083,390 filed Apr. 29, 1998; 60/085,224 filed May 13, 1998; 60/085,223 filed May 13, 1998; 60/085,222 filed May 13, 1998; 60/086,186 filed May 21, 1998; 60/086,187 filed May 21, 1998; 60/086,185 filed May 21, 1998; 60/086,184 filed May 21, 1998; 60/086,183 filed May 21, 1998; 60/086,188 filed May 21, 1998; 60/087,422 filed Jun. 1, 1998; 60/089,524 filed Jun. 16, 1998; 60/089,810 filed Jun. 18, 1998; 60/089,814 Jun. 18; 1998; 60/089,793 filed Jun. 18, 1998; 60/090,170 filed Jun. 22, 1998; 60/090,928 Jun. 26, 1998; 60/091,035 filed Jun. 29, 1998; 60/091,405 filed Jun. 30, 1998; 60/092,036 filed Jul. 8, 1998; 60/099,667 filed Sep. 9, 1998; 60/099,670 filed Sep. 9, 1998; 60/099,697 filed Sep. 9, 1998; 60/100,674 filed Sep. 16, 1998; 60/100,673 filed Sep. 16, 1998; 60/100,672 filed Sep. 16, 1998; 60/101,131 filed Sep. 21, 1998; 60/101,132 Sep. 21, 1998; 60/101,130 filed Sep. 21, 1998; 60/101,508 filed Sep. 22, 1998; 60/101,344 filed Sep. 22, 1998; 60/101,347 filed Sep. 22, 1998; 60/101,343 filed Sep. 22, 1998; 60/101,707 filed Sep. 25, 1998; 60/104,126 filed Oct. 13, 1998; 60/104,128 filed Oct. 13, 1998; 60/104,127 filed Oct. 13, 1998; 60/104,124 filed Oct. 13, 1998; 60/104,123 filed Oct. 13, 1998; 60/109,018 filed Nov. 19, 1998; 60/108,996 filed Nov. 18, 1998; 60/111,981 filed Dec. 11, 1998; and 60/113,224 filed Dec. 22, 1998; and claims the benefit under 35 U.S.C. § 120 as a continuation-in-part application of U.S. application Ser. No. 09/199,129 filed Nov. 24, 1998 now abandoned; Ser. No. 09/210,297 filed Dec. 8, 1998 now abandoned; and Ser. No. 09/229,413 filed Jan. 12, 1999 now abandoned, the disclosures of which applications are herein incorporated by reference in their entirety.

FIELD OF THE INVENTION

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean plants associated with the sucrose pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

BACKGROUND OF THE INVENTION

Carbon fixed during photosynthesis is either retained in the chloroplast and converted to a storage carbohydrate, for example, starch, or it is transferred to the cytosol in the form of triose phosphates and converted to sucrose. The newly synthesized sucrose in source tissues is a major transported form of reduced carbon in higher plants and can be either metabolized into other carbohydrates, stored in the vacuole or exported to other plant tissues. Plant tissues where sucrose is synthesized, such as leaves, are often referred to as ‘source’ tissues. Translocated sucrose is retained in ‘sink’ tissues (such as expanding leaves, growing seeds, flowers, roots or tubers, and fruit) and may be assimilated, or further metabolized to sustain cell maintenance or fuel growth, or be converted to alternative storage compounds (e.g., starch, fats). The relative type and size of these carbohydrate pools vary during tissue development, between different plant species, and within the same species subject to different environmental conditions. Such differences are reported to affect the yield and quality of agricultural produce.

Sucrose synthesis and catabolism are reported to be highly coordinated and regulated processes that may also be coordinately regulated with other dedicated metabolic pathways in a particular plant, plant organ or cell type. Sucrose synthesis is reported to be coordinately regulated with starch metabolism and photosynthesis in green ‘source’ plant tissues. Sucrose supply by transport mechanisms to actively growing ‘sink’ tissues is reported to be coordinated with plant development. In growing sink tissues, the supply of carbohydrate is reported to be important to other metabolic pathways and physiological processes including respiration, starch biosynthesis, cell wall biogenesis, lipid and protein biosynthesis. Sucrose synthesis and/or transport is also reported to play a role in the carbohydrate capacity that is available to growing fruits and seeds. Sucrose resynthesis during seed germination is reported to play a role in seedling vigor and agronomic stand establishment in many plant species during early plant development.

In many plant species, enzymes of pathways involved in sucrose metabolism can play a role in plant physiology and plant growth and development. Compartmentation and temporal regulation of genes and enzymes of sucrose metabolic pathways can allow multiple pathways to utilize sucrose as a common metabolite. Flux through a particular sucrose metabolic pathway can define the utilization of sucrose in any tissue or developmental stage. Sucrose and its metabolite products have been reported to play a role in gene regulation and expression of the sucrose pathway and other metabolic pathways in plants.

Reviews on sucrose metabolism in plants include Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Hawker, In: Biochemistry of Storage Carbohydrates in Green Plants, Dey and Dixon, eds., Academic Press, London, 1-51 (1985); Huber et al., In: Carbon Partitioning Within and Between Organisms, Pollock et al., eds., Bios Scientific, Oxford, 1-26 (1992); Stitt et al., In: Biochemistry of Plants, Vol 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987); Quick and Schaffer, In: Photoassimilate Distribution In: Plants And Crops, Zamski and Schaffer, eds., Marcel Dekker Inc., New York, 115-156 (1996), all of which are herein incorporated by reference in their entirety.

The synthesis of sucrose precursors (triose and hexose phosphates) is derived from either photosynthetic CO₂ fixation or degradation of previously deposited storage reserves. One substrate for sucrose synthesis in photosynthetic tissues is three carbon sugar phosphates. These are exported from the chloroplast during photosynthesis, predominantly in the form of triose phosphates. The pool of triose phosphates, dihydroxyacetone phosphate (“DHAP”), and glyceraldehyde-3-phosphate (“GAP”), is maintained at equilibrium within the cytoplasm by triose phosphate isomerase (EC 5.3.1.1). A subsequent reaction involves an aldol condensation of DHAP and GAP, catalyzed by the enzyme fructose 1,6-bisphosphate aldolase (often called aldolase) (EC 4.1.2.13) to form fructose 1,6-bisphosphate (“F1,6BP”). Fructose-1,6-bisphosphatase (“FBPase”) (EC 3.1.3.11) catalyzes the cleavage of phosphate from the C1 carbon of fructose-1,6-bisphosphate to form fructose-6-phosphate (“F6P”). This reaction is essentially irreversible and has been reported to represent the first committed step within the pathway of sucrose synthesis. The cytosolic FBPase has been reported to be subject to allosteric regulation and may serve to coordinate the rate of sucrose synthesis with that of photosynthesis. Fructose 2,6-bisphosphate (“F2,6BP”) is reported to be a regulator of FBPase (Black et al., In: Regulation of Carbohydrate Partitioning In Photosynthetic Tissue, Heath and Preiss, eds., Waverly, Baltimore, 109-126 (1985); Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987), both of which are herein incorporated by reference in their entirety). The concentration of F2,6BP is reported to be controlled in plants by two enzymes, fructose-2,6-bisphosphatase (F2,6Bpase) (EC 3.1.3.46) and fructose-6-phosphate,2-kinase (F6P,2K) (EC 2.7.1.105) (Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 153-181 (1990), the entirety of which is herein incorporated by reference).

Glucose-6-phosphate (“G6P”) and glucose-1-phosphate (“G1P”) are reported to be maintained in equilibrium with the F6P pool by the action of phosphoglucoisomerase (“PGI”) (EC 5.3.1.9) and phosphoglucomutase (“PGM”) (EC 5.4.2.2), respectively. Uridine diphosphate glucose (“UDPG”) and pyrophosphate (“PPi”) are formed from uridine triphosphate (“UTP”) and G1P catalyzed by the enzyme UDPG-pyrophosphorylase (“UDPGase”) (EC 2.7.7.9). This reaction is reversible and net flux in the direction of sucrose synthesis is reported to require removal of its products, particularly PPi. A pyrophosphate-dependent proton pump, vacuolar H⁺-translocating-pyrophosphatase (EC 3.6.1.1), has been identified within the vacuolar membrane and has been reported to utilize pyrophosphate to sustain a proton gradient formed between these two compartments (Rea et al., Trends in Biol. Sci. 17: 348-353 (1992), the entirety of which is herein incorporated by reference).

A pyrophosphate-dependent fructose-6-phosphate phosphotransferase (“PFP”) (EC 2.7.1.90) is also present in the cytoplasm and catalyzes the reversible production of F1,6BP and Pi from F6P and PPi. One reported function of PFP is to operate in a futile cycle with the cytosolic FBPase, and function as a “pseudopyrophosphatase” recycling PPi. Uridine diphosphate glucose is then combined with F6P to form sucrose-6-phosphate (“S6P”). This reaction is catalyzed by sucrose phosphate synthase (“SPS”) (EC 2.4.1.14). Attachment of UDP to the glucose moiety activates the C1 carbon atom of UDPG, which is necessary for the subsequent formation of a glycosidic bond in sucrose. In certain organisms, SPS is capable of using adenine diphosphate glucose (“ADPG”), instead of UDPG, as a substrate. The use of nucleotide biphosphate sugars is a feature of metabolic pathways leading to the production of disaccharides and polysaccharides. SPS is reported to be subject to allosteric and covalent regulation and, in conjunction with the cytosolic FBPase, reportedly serves to coordinate the rate of sucrose synthesis with the rate of photosynthesis. The reported final reaction in the pathway is catalyzed by sucrose-6-phosphate phosphatase (“SPPase” or “SPP”) (EC 3.1.3.24), which catalyzes the hydrolysis of S6P to sucrose. It has been reported that SPS and SPPase may associate to form a multienzyme complex, that the rate of sucrose-6-phosphate synthesis by SPS is enhanced in the presence of SPP, and that the rate of sucrose-6-phosphate hydrolysis by SPP is increased in the presence of SPS (Echeverria et al., Plant Physiol. 115: 223-227 (1997), herein incorporated by reference in its entirety).

I. Sucrose Synthesis

Reviews describing fructose-1,6-bisphosphatase (“FBPase”, EC 3.1.3.11) include those by Hers and Van Shaftingen, Biochem J. 206:1-12 (1982), the entirety of which is herein incorporated by reference, and Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41:153-181 (1990). Two isoforms of FBPase are reported to exist in plants. The first isoform is associated with the plastid and occurs largely in photosynthetic plastids. The second isoform, located in the cytoplasm, is reported to be involved in both gluconeogenesis and sucrose synthesis (Zimmerman et al., J. Biol. Chem. 253: 5952-5956 (1978); Stitt and Heldt, Planta 164: 179-188 (1985), both of which are hereby incorporated by reference in their entirety). FBPase catalyzes an irreversible reaction in the direction of F6P synthesis in vivo and has been reported to represent the first committed step in the pathway of sucrose synthesis. The properties of the enzyme are reported to involve the action of several regulatory metabolites (Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987)). The enzyme reportedly has a high affinity for its substrate F1,6BP, a requirement for Mg²⁺, a requirement for a neutral pH, is weakly inhibited (Km 2-4 μm) by adenosine monophosphate (AMP), and is strongly inhibited by the regulatory metabolite F2,6BP (Hers and Van Shaftingen, Biochem J. 206: 1-12 (1982); Black et al., In: Regulation of Carbohydrate Partitioning In Photosynthetic Tissue, Heath and Preiss, eds., Waverly, Baltimore, 109-126 (1985); Huber, Annu. Rev. Plant Physiol. 37: 233-246 (1986); Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987), all of which are herein incorporated by reference in their entirety). F2,6BP is also an activator of PFP and reportedly plays a role in the regulation of gluconeogenetic and respiratory metabolism.

The concentration of F2,6BP is reportedly determined in plants by two enzymes, fructose-2,6-bisphosphatase (“F2,6BPase”) (EC 3.1.3.46) and fructose-6-phosphate,2-kinase (“F6P,2K”) (EC 2.7.1.105). A review of these enzymes is provided by Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 153-181 (1990). Regulation of the activity of the F1,6FBPase and the rate of sucrose synthesis is reported to be, at least in part, brought about by changes in the concentration of F2,6BP.

Sucrose phosphate synthase (SPS (EC 2.4.1.14)) catalyzes a reaction that is displaced from equilibrium in vivo in the direction of S6P synthesis and is reported as an essentially irreversible reaction in vivo (Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987); Lunn and Rees, Biochem. J. 267: 739-743 (1990), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,665,892, the entirety of which is herein incorporated by reference). SPS has been purified from spinach and Zea mays, and the amino acid and cDNA sequences have been published (Worrel et al., Plant Cell 3:1121-1130 (1991); Klein et al., Planta 190: 498-510 (1993); Sonnewald et al., Planta 189: 174-181 (1993), all of which are herein incorporated by reference in their entirety). The enzyme has a subunit molecular weight of 117 kDa from spinach (Klein et al., Planta 190: 498-510 (1993); Sonnewald et al., Planta 189: 174-181 (1993), both of which are herein incorporated by reference) and pea (Lunn and Rees, Phytochem. 29: 1057-1063 (1990), the entirety of which is herein incorporated by reference) and 135 kDa from Zea mays (Worrel et al., Plant Cell 3:1121-1130 (1991)). The native enzyme reportedly exists as a tetramer (Walker and Huber, Plant Physiol. 89: 518-524 (1988); Lunn and Rees, Phytochem. 29: 1057-1063 (1990); Worrel et al., Plant Cell 3:1121-1130 (1991), although dimeric molecular weights have been reported (Klein et al., Planta 190: 498-510 (1993), the entirety of which is herein incorporated by reference). Activity has been observed for SPS at both dimeric and tetrameric molecular weights (Sonnewald et al., Planta 189:174-181 (1993), the entirety of which is herein incorporated by reference).

SPS is located in the cytosol, has a neutral pH optimum, and has been detected in all plant tissues which undertake active sucrose synthesis. SPS is also reported to undertake active sucrose synthesis. An increase in abundance of the enzyme is has been reported during the development of leaves, germination of seeds and ripening of fruit. The enzyme has been reported to be subject to regulation by metabolites and is activated by G6P and is inhibited by Pi. Pi and GP6 are reported to act competitively at an allosteric site of the enzyme. In the presence of high Pi concentrations, the enzyme is phosphorylated which reduces activity of the enzyme. It has also been reported that light-induced photosynthesis increases the activity of SPS in crude extracts (Sicher and Kremer, Plant Physiol. 79: 910-912 (1984), Sicher and Kremer, Plant Physiol. 79: 695-698 (1985); Pollock and Housley, Ann. Bot. 55: 593-596 (1985), all of which are herein incorporated by reference in their entirety). In addition, it has been reported that compounds altering the phosphate status of the leaf can simulate the effects of light. Feeding leaves mannose, which sequesters phosphate by its conversion to the non-metabolized mannose-6-P, has been reported to cause activation of SPS (Stitt et al., Planta 174: 217-230 (1988), the entirety of which is herein incorporated by reference).

The phosphorylation and dephosphorylation of SPS is catalyzed by SPS-phosphatase and SPS-kinase, respectively (Huber et al., Plant Physiol. 99: 1275-1278 (1992). Hydrolysis of sucrose-6-P to sucrose is catalyzed by sucrose-6-phosphatase (SPPase or SPP) (EC 3.1.3.24). The activity of both SPS and SPP is reported to be affected by a multienzyme complex between SPS and SPP (Echeverria et al., Plant Physiol. 115: 223-227 (1997)).

Regulatory properties of SPS and FBPase are reported to coordinate the rate of sucrose synthesis with that of photosynthesis (Stitt, In: Plant Physiology, Biochemistry and Molecular Biology, Dennis and Turpin, eds., Singapore, London, 319-340 (1990), the entirety of which is herein incorporated by reference). When photosynthesis produces triose phosphate in excess of the rate of sucrose synthesis, a feed-forward activation of sucrose synthesis occurs. Triose phosphate crosses the chloroplast membrane in exchange for cytosolic Pi. Under these conditions, F6P,2-kinase activity is reduced and the inhibition of F2,6Bpase is decreased.

As cytosolic F2,6BP falls, F2,6BPase activity increases, and F6P levels increase. Hexose phosphate levels are reported to increase due to PGM and PGI, and with low Pi, activate SPS and F1,6BPase. Reduction in rate of photosynthesis must result in a deactivation of sucrose synthesis, which occurs through decreased cytosolic triose-P, increased Pi and ultimately increased F2,6BP concentration and reduced SPS activity (Stitt, Phil. Trans. R Soc. Lond. B 342: 225-233 (1993); Huber et al., Plant Physiol. 99: 1275-1278 (1992); Neuhaus et al., Planta 181: 583-592 (1990), both of which are herein incorporated by reference).

II. Metabolic Pathways of Sucrose Catabolism

Sucrose can initially be cleaved by invertases (EC 3.2.1.26) or by sucrose synthases (EC 2.4.1.13). Invertases, which are classified as acid or alkaline in pH preference (Karuppiah et al., Plant Physiol. 91: 993-998 (1989); Fahrendorf and Beck, Planta 180: 237-244 (1990); Iwatsubo et al., Biosc. Biotech. Biochem. 56: 1959-1962 (1992); Unger et al., Plant Physiol. 104: 1351-1357 (1994); Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982), all of which are herein incorporated by reference in their entirety), irreversibly cleave sucrose into glucose and fructose, both of which is usually phosphorylated for further metabolism. The invertase pathway usually is associated with rapidly growing sink tissues such as expanding leaves, expanding internodes, flower petals, and early fruit development (Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Huber, Plant Physiol. 91: 656-662 (1989); Morris and Arthur, Phytochem. 23: 2163-2167 (1984); Hawker et al., Phytochem. 15: 1441-1443 (1976); Schaffer et al., Plant Physiol. 69: 151-155 (1987), all of which are herein incorporated by reference in their entirety).

Sucrose synthase carries out the kinetically reversible transglycosylation of sucrose and UDP into fructose and UDPG, requiring only the phosphorylation of fructose for additional metabolism. Polysaccharide biosynthesis in sink tissues may utilize a sucrose synthase mediated sucrose catabolism (Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Dale and Housley Plant Physiol. 82: 7-10 (1986), all of which are herein incorporated by reference). Respiring tissues reportedly utilize either sucrose synthase or invertase metabolic pathways (Echeverria and Humphreys, Phytochem. 23: 2173-2178 (1984); Uritani and Asahi, In: The Biochemistry of Plants Vol. 2, Davies, ed., Academic Press, New York, 463-487 (1980), all of which are herein incorporated by reference in their entirety). Tissues that are undergoing respiration, starch biosynthesis, amino acid and fatty acid synthesis, rapid expansion or growth, and other cellular metabolism, can utilize several sucrose metabolic pathways which may be temporally or compartmentally regulated (Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Doehlert, Plant Physiol. 78: 560-567 (1990); Doehlert and Choury, In: Recent Advances in Phloem Transport and Assimilate Compartmentation, Bonnemain et al., eds., Ouest editions, Nantes, France, 187-195 (1991); Delmer and Stone, In: The Biochemistry of Plants, Vol. 14, Preiss, ed., Academic Press, San Diego, 373-420 (1988); Maas et al., EMBO J. 9: 3447-3452 (1990), all of which are herein incorporated by reference in their entirety).

Hexose kinases are a class of enzymes responsible for the phosphorylation of hexoses, and are classified into two groups. Hexokinase (EC 2.7.1.1) can phosphorylate either glucose or fructose, with different isoforms often unique to different tissues or plant species. Different isoforms can have affinities for different hexoses (Turner and Copeland, Plant Physiol. 68: 1123-1127 (1981), the entirety of which is herein incorporated by reference; Copeland and Turner, In: The Biochemistry of Plants, Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987), the entirety of which is herein incorporated by reference). Hexokinases include fructokinases (EC 2.7.1.11), which typically have specific affinities for fructose (Doehlert, Plant Physiol. 89: 1042-1048 (1989); Renz and Stitt Planta 190: 166-175 (1993), both of which are herein incorporated by reference). Fructokinases can also be specific in their affinity for nucleotides. The extent to which a fructokinase utilizes UTP may play a physiological role in how efficiently UDP can be recycled for sucrose synthase activity in a particular tissue (Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Xu et al., Plant Physiol. 90: 635-642 (1989), both of which are herein incorporated by reference). UDP levels for the sucrose synthase reaction may be maintained, even in the case of an ATP-specific fructokinase, by the enzyme NDP-kinase (EC 2.7.4.6).

NDP-kinase has been reported in several plant tissues (Kirkland and Turner, J. Biochem. 72: 716-720 (1959); Bryce and Nelson, Plant Physiol. 63: 312-317 (1979); Dancer et al., Plant Physiol. 92: 637-641 (1990); Yano et al., Plant Molec. Biol. 23: 1087-1090 (1993), all of which are herein incorporated by reference in their entirety). Fructokinase can be substrate inhibited by fructose. In addition, sucrose synthase can be inhibited by fructose (Doehlert, Plant Sci. 52: 153-157 (1987); Morell and Copeland, Plant Physiol. 78: 140-154 (1985), Ross and Davies, Plant Physiol. 100: 1008-1013 (1992), all of which are herein incorporated by reference in their entirety). Whereas plant tissues where sucrose is catabolized by sucrose synthase predominantly contain fructokinases (Xu et al., Plant Physiol. 90: 635-642 (1989); Kursanov et al., Soviet Plant Physiol. 37: 507-515 (1990); Ross et al., Plant Physiol. 90: 748-756 (1994)), plant tissues where sucrose is catabolized by invertase often contain hexokinases (Nakamura et al., Plant Physiol. 81: 215-220 (1991)). Tissues which have both invertase and sucrose synthase activity may contain both hexose kinases (Nakamura et al., Plant Physiol. 81: 215-220 (1991), the entirety of which is herein incorporated by reference). F6P resulting from hexose kinase activity can be further metabolized in glycolysis or used in resynthesis of sucrose by SPS. G6P resulting from hexose kinase activity can enter the pentose phosphate pathway, via G6P dehydrogenase (EC 1.1.1.49), or be converted to F6P by phosphoglucoisomerase (“PGI”) (EC 5.3.1.9) or G1P by phosphoglucomutase (“PGM”) (EC 5.4.2.2) (Rees, In: Encyclopedia of Plant Physiology Vol 18, Douce and Day, eds., Springer Verlag, Berlin, 391-417 (1985); Copeland and Turner, In: The Biochemistry of Plants Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987); Foster and Smith, Planta 180: 237-244 (1993), all of which are herein incorporated by reference in their entirety).

PGI and PGM are reported to be ubiquitous and reversible with commitments of G6P to either F6P or G1P resulting from fluxes in metabolites further along each pathway, i.e., depending on the cell needs for glycolysis (F6P) or starch biosynthesis (G1P) (Edwards and Rees, Phytochem. 25: 2033-2039 (1986); Kursanov et al., Soviet Plant Physiol. 37: 507-515 (1990); Tobias et al., Plant Physiol. 99: 140-145 (1992), all of which are herein incorporated by reference in their entirety). UDPG formed by sucrose synthase may be utilized directly for cellulose or callose biosynthesis via UDP-glucose dehydrogenase (EC 1.1.1.2) (Robertson et al., Phytochem. 39: 21-28 (1995), the entirety of which is herein incorporated by reference), can be used for sucrose synthesis by SPS or sucrose synthase, or for glycolysis or starch metabolism dependent on further metabolism by UDP-glucose pyrophosphorylase (EC 2.7.7.9). UDP-glucose phosphorylase has been reported to be a largely reversible enzyme (Kleczkowski, Phytochem. 37: 1507-1515 (1994), the entirety of which is herein incorporated by reference). Flux through UDP-glucose pyrophosphorylase is reported to be influenced by metabolite levels and utilization of reaction products further along in the pathways (Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Zrenner et al., Planta 190: 247-252 (1993), all of which are herein incorporated by reference in their entirety). The reversibility of PGI, PGM and UDPGPPase has been reported to provide for metabolic variability and networking in metabolism, independent of which initial enzyme cleaved sucrose.

The fate of F6P reportedly plays a role in carbohydrate metabolism. NTP-phosphofructokinase (PFK) (EC 2.7.1.11) (Copeland and Turner, In: The Biochemistry of Plants Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987); Dennis and Greyson, Plant Physiol. 69: 395-404 (1987); Rees, In: The Biochemistry of Plants Vol. 14, Preiss, ed., Academic Press, San Diego, 1-33 (1988), all of which are herein incorporated by reference in their entirety) is reported to irreversibly convert F6P to F16BP and is associated with glycolysis. The reverse reaction of F16BP to F6P, associated with gluconeogenesis, is essentially irreversible, and is catalyzed by FBPase (EC 3.1.3.11) (Black et al., Plant Physiol. 69: 387-394 (1987). Both reactions may be carried out in a reversible manner by a PPi-dependent fructose-6-phosphate phosphotransferase or PPi-phosphofructokinase (PFP; EC 2.7.1.90) (Black et al., Plant Physiol. 69: 387-394 (1987).

PPi-dependent fructose-6-phosphate phosphotransferase or PPi-phosphofructokinase is reported to play a role in the generation of biosynthetic intermediates (Dennis and Greyson, Plant Physiol. 69: 395-404 (1987); Tobias et al., Plant Physiol. 99: 146-152 (1992), the entirety of which is herein incorporated by reference) in addition to the cycling of PPi for UDPGPPase and ultimately UDP for sucrose synthase (Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Black et al., Plant Physiol. 69: 387-394 (1987); Rees, In: The Biochemistry of Plants Vol. 14, Preiss, ed., Academic Press, San Diego, 1-33 (1988), all of which are herein incorporated by reference in their entirety).

II. Expressed Sequence TAG Nucleic Acid Molecules

Expressed sequence tags, or ESTs are randomly sequenced members of a cDNA library (or complementary DNA)(McCombie et al., Nature Genetics 1:124-130 (1992); Kurata et al., Nature Genetics 8:365-372 (1994); Okubo et al., Nature Genetics 2:173-179 (1992), all of which references are incorporated herein in their entirety). The randomly selected clones comprise insets that can represent a copy of up to the full length of a mRNA transcript.

Using conventional methodologies, cDNA libraries can be constructed from the mRNA (messenger RNA) of a given tissue or organism using poly dT primers and reverse transcriptase (Efstratiadis et al., Cell 7:279-3680 (1976), the entirety of which is herein incorporated by reference; Higuchi et al., Proc. Natl. Acad. Sci. (U.S.A.) 73:3146-3150 (1976), the entirety of which is herein incorporated by reference; Maniatis et al., Cell 8:163-182 (1976) the entirety of which is herein incorporated by reference; Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference; Okayama et al., Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference; Gubler et al., Gene 25:263-269 (1983), the entirety of which is herein incorporated by reference).

Several methods may be employed to obtain full-length cDNA constructs. For example, terminal transferase can be used to add homopolymeric tails of dC residues to the free 3′ hydroxyl groups (Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference). This tail can then be hybridized by a poly dG oligo which can act as a primer for the synthesis of full length second strand cDNA. Okayama and Berg, Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference, report a method for obtaining full length cDNA constructs. This method has been simplified by using synthetic primer-adapters that have both homopolymeric tails for priming the synthesis of the first and second strands and restriction sites for cloning into plasmids (Coleclough et al., Gene 34:305-314 (1985), the entirety of which is herein incorporated by reference) and bacteriophage vectors (Krawinkel et al., Nucleic Acids Res. 14:1913 (1986), the entirety of which is herein incorporated by reference; Han et al., Nucleic Acids Res. 15:6304 (1987), the entirety of which is herein incorporated by reference).

These strategies have been coupled with additional strategies for isolating rare mRNA populations. For example, a typical mammalian cell contains between 10,000 and 30,000 different mRNA sequences (Davidson, Gene Activity in Early Development, 2nd ed., Academic Press, New York (1976), the entirety of which is herein incorporated by reference). The number of clones required to achieve a given probability that a low-abundance mRNA will be present in a cDNA library is N=(1n(1−P))/(1n(1−1/n)) where N is the number of clones required, P is the probability desired and 1/n is the fractional proportion of the total mRNA that is represented by a single rare mRNA (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press (1989), the entirety of which is herein incorporated by reference).

A method to enrich preparations of mRNA for sequences of interest is to fractionate by size. One such method is to fractionate by electrophoresis through an agarose gel (Pennica et al., Nature 301:214-221 (1983), the entirety of which is herein incorporated by reference). Another such method employs sucrose gradient centrifugation in the presence of an agent, such as methylmercuric hydroxide, that denatures secondary structure in RNA (Schweinfest et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:4997-5000 (1982), the entirety of which is herein incorporated by reference).

A frequently adopted method is to construct equalized or normalized cDNA libraries (Ko, Nucleic Acids Res. 18:5705-5711 (1990), the entirety of which is herein incorporated by reference; Patanjali et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:1943-1947 (1991), the entirety of which is herein incorporated by reference). Typically, the cDNA population is normalized by subtractive hybridization (Schmid et al., J. Neurochem. 48:307-312 (1987), the entirety of which is herein incorporated by reference; Fargnoli et al., Anal. Biochem. 187:364-373 (1990), the entirety of which is herein incorporated by reference; Travis et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1696-1700 (1988), the entirety of which is herein incorporated by reference; Kato, Eur. J. Neurosci. 2:704-711 (1990); and Schweinfest et al., Genet. Anal. Tech. Appl. 7:64-70 (1990), the entirety of which is herein incorporated by reference). Subtraction represents another method for reducing the population of certain sequences in the cDNA library (Swaroop et al., Nucleic Acids Res. 19:1954 (1991), the entirety of which is herein incorporated by reference).

ESTs can be sequenced by a number of methods. Two basic methods may be used for DNA sequencing, the chain termination method of Sanger et al., Proc. Natl. Acad. Sci. (U.S.A.) 74:5463-5467 (1977), the entirety of which is herein incorporated by reference and the chemical degradation method of Maxam and Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564 (1977), the entirety of which is herein incorporated by reference. Automation and advances in technology such as the replacement of radioisotopes with fluorescence-based sequencing have reduced the effort required to sequence DNA (Craxton, Methods 2:20-26 (1991), the entirety of which is herein incorporated by reference; Ju et al., Proc. Natl. Acad. Sci. (U.S.A.) 92:4347-4351 (1995), the entirety of which is herein incorporated by reference; Tabor and Richardson, Proc. Natl. Acad. Sci. (U.S.A.) 92:6339-6343 (1995), the entirety of which is herein incorporated by reference). Automated sequencers are available from, for example, Pharmacia Biotech, Inc., Piscataway, N.J. (Pharmacia ALF), LI-COR, Inc., Lincoln, Neb. (LI-COR 4,000) and Millipore, Bedford, Mass. (Millipore BaseStation).

In addition, advances in capillary gel electrophoresis have also reduced the effort required to sequence DNA and such advances provide a rapid high resolution approach for sequencing DNA samples (Swerdlow and Gesteland, Nucleic Acids Res. 18:1415-1419 (1990); Smith, Nature 349:812-813 (1991); Luckey et al., Methods Enzymol. 218:154-172 (1993); Lu et al., J. Chromatog. A. 680:497-501 (1994); Carson et al., Anal. Chem. 65:3219-3226 (1993); Huang et al., Anal. Chem. 64:2149-2154 (1992); Kheterpal et al., Electrophoresis 17:1852-1859 (1996); Quesada and Zhang, Electrophoresis 17:1841-1851 (1996); Baba, Yakugaku Zasshi 117:265-281 (1997), all of which are herein incorporated by reference in their entirety).

ESTs longer than 150 nucleotides have been found to be useful for similarity searches and mapping (Adams et al., Science 252:1651-1656 (1991), herein incorporated by reference). ESTs, which can represent copies of up to the full length transcript, may be partially or completely sequenced. Between 150-450 nucleotides of sequence information is usually generated as this is the length of sequence information that is routinely and reliably produced using single run sequence data. Typically, only single run sequence data is obtained from the cDNA library (Adams et al., Science 252:1651-1656 (1991). Automated single run sequencing typically results in an approximately 2-3% error or base ambiguity rate (Boguski et al., Nature Genetics 4:332-333 (1993), the entirety of which is herein incorporated by reference).

EST databases have been constructed or partially constructed from, for example, C. elegans (McCombrie et al., Nature Genetics 1:124-131 (1992)), human liver cell line HepG2 (Okubo et al., Nature Genetics 2:173-179 (1992)), human brain RNA (Adams et al., Science 252:1651-1656 (1991); Adams et al., Nature 355:632-635 (1992)), Arabidopsis, (Newman et al., Plant Physiol. 106:1241-1255 (1994)); and rice (Kurata et al., Nature Genetics 8:365-372 (1994)).

III. Sequence Comparisons

A characteristic feature of a DNA sequence is that it can be compared with other DNA sequences. Sequence comparisons can be undertaken by determining the similarity of the test or query sequence with sequences in publicly available or proprietary databases (“similarity analysis”) or by searching for certain motifs (“intrinsic sequence analysis”)(e.g. cis elements)(Coulson, Trends in Biotechnology 12:76-80 (1994), the entirety of which is herein incorporated by reference); Birren et al., Genome Analysis 1: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997), the entirety of which is herein incorporated by reference).

Similarity analysis includes database search and alignment. Examples of public databases include the DNA Database of Japan (DDBJ) (available on the worldwide web at ddbj.nig.ac.jp); Genebank (available on the worldwide web at the ncbi website at: /Web/Search/Index.html); and the European Molecular Biology Laboratory Nucleic Acid Sequence Database (EMBL) (available on the worldwide web at ebi.ac.uk/ebi_docs/embl_db/embl-db.html). Other appropriate databases include dbEST (available on the worldwide web at the ncbi website at:/dbEST /index.html), SwisProt (available on the worldwide web at ebi.ac.uk/ebi_docs/swisprot_db/swisshome.html), PIR (available on the worldwide web at nbrt.georgetown.edu/pir), and The Institute for Genome Research (available on the worldwide web at tigr.org/tdb/tdb.html).

A number of different search algorithms have been developed, one example of which are the suite of programs referred to as BLAST programs. There are five implementations of BLAST, three designed for nucleotide sequences queries (BLASTN, BLASTX and TBLASTX) and two designed for protein sequence queries (BLASTP and TBLASTN) (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al, Genome Analysis 1, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997)).

BLASTN takes a nucleotide sequence (the query sequence) and its reverse complement and searches them against a nucleotide sequence database. BLASTN was designed for speed, not maximum sensitivity and may not find distantly related coding sequences. BLASTX takes a nucleotide sequence, translates it in three forward reading frames and three reverse complement reading frames and then compares the six translations against a protein sequence database. BLASTX is useful for sensitive analysis of preliminary (single-pass) sequence data and is tolerant of sequencing errors (Gish and States, Nature Genetics 3:266-272 (1993), the entirety of which is herein incorporated by reference). BLASTN and BLASTX may be used in concert for analyzing EST data (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1:543-559 (1997)).

Given a coding nucleotide sequence and the protein it encodes, it is often preferable to use the protein as the query sequence to search a database because of the greatly increased sensitivity to detect more subtle relationships. This is due to the larger alphabet of proteins (20 amino acids) compared with the alphabet of nucleic acid sequences (4 bases), where it is far easier to obtain a match by chance. In addition, with nucleotide alignments, only a match (positive score) or a mismatch (negative score) is obtained, but with proteins, the presence of conservative amino acid substitutions can be taken into account. Here, a mismatch may yield a positive score if the non-identical residue has physical/chemical properties similar to the one it replaced. Various scoring matrices are used to supply the substitution scores of all possible amino acid pairs. A general purpose scoring system is the BLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61 (1993), the entirety of which is herein incorporated by reference), which is currently the default choice for BLAST programs. BLOSUM62 is tailored for alignments of moderately diverged sequences and thus may not yield the best results under all conditions. Altschul, J. Mol. Biol. 36:290-300 (1993), the entirety of which is herein incorporated by reference, describes a combination of three matrices to cover all contingencies. This may improve sensitivity, but at the expense of slower searches. In practice, a single BLOSUM62 matrix is often used but others (PAM40 and PAM250) may be attempted when additional analysis is necessary. Low PAM matrices are directed at detecting very strong but localized sequence similarities, whereas high PAM matrices are directed at detecting long but weak alignments between very distantly related sequences.

Homologues in other organisms are available that can be used for comparative sequence analysis. Multiple alignments are performed to study similarities and differences in a group of related sequences. CLUSTAL W is a multiple sequence alignment package that performs progressive multiple sequence alignments based on the method of Feng and Doolittle, J. Mol. Evol. 25:351-360 (1987), the entirety of which is herein incorporated by reference. Each pair of sequences is aligned and the distance between each pair is calculated; from this distance matrix, a guide tree is calculated and all of the sequences are progressively aligned based on this tree. A feature of the program is its sensitivity to the effect of gaps on the alignment; gap penalties are varied to encourage the insertion of gaps in probable loop regions instead of in the middle of structured regions. Users can specify gap penalties, choose between a number of scoring matrices, or supply their own scoring matrix for both pairwise alignments and multiple alignments. CLUSTAL W for UNIX and VMS systems is available by anonymous ftp at: ebi.ac.uk. Another program is MACAW (Schuler et al., Proteins Struct. Func. Genet. 9:180-190 (1991), the entirety of which is herein incorporated by reference, for which both Macintosh and Microsoft Windows versions are available. MACAW uses a graphical interface, provides a choice of several alignment algorithms and is available by anonymous ftp at the ncbi website at: nlm.nih.gov (directory/pub/macaw).

Sequence motifs are derived from multiple alignments and can be used to examine individual sequences or an entire database for subtle patterns. With motifs, it is sometimes possible to detect distant relationships that may not be demonstrable based on comparisons of primary sequences alone. Currently, the largest collection of sequence motifs in the world is PROSITE (Bairoch and Bucher, Nucleic Acid Research 22:3583-3589 (1994), the entirety of which is herein incorporated by reference). PROSITE may be accessed via either the ExPASy server on the World Wide Web or anonymous ftp site. Many commercial sequence analysis packages also provide search programs that use PROSITE data.

A resource for searching protein motifs is the BLOCKS E-mail server developed by Henikoff, Trends Biochem Sci. 18:267-268 (1993), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Nucleic Acid Research 19:6565-6572 (1991), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Proteins 17:49-61 (1993). BLOCKS searches a protein or nucleotide sequence against a database of protein motifs or “blocks.” Blocks are defined as short, ungapped multiple alignments that represent highly conserved protein patterns. The blocks themselves are derived from entries in PROSITE as well as other sources. Either a protein query or a nucleotide query can be submitted to the BLOCKS server; if a nucleotide sequence is submitted, the sequence is translated in all six reading frames and motifs are sought for these conceptual translations. Once the search is completed, the server will return a ranked list of significant matches, along with an alignment of the query sequence to the matched BLOCKS entries.

Conserved protein domains can be represented by two-dimensional matrices, which measure either the frequency or probability of the occurrences of each amino acid residue and deletions or insertions in each position of the domain. This type of model, when used to search against protein databases, is sensitive and usually yields more accurate results than simple motif searches. Two popular implementations of this approach are profile searches such as GCG program ProfileSearch and Hidden Markov Models (HMMs)(Krough et al., J. Mol. Biol. 235:1501-1531, (1994); Eddy, Current Opinion in Structural Biology 6:361-365, (1996), both of which are herein incorporated by reference in their entirety). In both cases, a large number of common protein domains have been converted into profiles, as present in the PROSITE library, or HHM models, as in the Pfam protein domain library (Sonnhammer et al., Proteins 28:405-420 (1997), the entirety of which is herein incorporated by reference). Pfam contains more than 500 HMM models for enzymes, transcription factors, signal transduction molecules and structural proteins. Protein databases can be queried with these profiles or HMM models, which will identify proteins containing the domain of interest. For example, HMMSW or HMMFS, two programs in a public domain package called HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can be used.

PROSITE and BLOCKS represent collected families of protein motifs. Thus, searching these databases entails submitting a single sequence to determine whether or not that sequence is similar to the members of an established family. Programs working in the opposite direction compare a collection of sequences with individual entries in the protein databases. An example of such a program is the Motif Search Tool, or MoST (Tatusov et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:12091-12095 (1994), the entirety of which is herein incorporated by reference). On the basis of an aligned set of input sequences, a weight matrix is calculated by using one of four methods (selected by the user). A weight matrix is simply a representation, position by position of how likely a particular amino acid will appear. The calculated weight matrix is then used to search the databases. To increase sensitivity, newly found sequences are added to the original data set, the weight matrix is recalculated and the search is performed again. This procedure continues until no new sequences are found.

SUMMARY OF THE INVENTION

The present invention provides a substantially purified nucleic acid molecule that encodes a maize or a soybean enzyme or fragment thereof, wherein the maize or the soybean enzyme is selected from the group consisting of: (a) triose phosphate isomerase; (b) fructose 1,6-bisphosphate aldolase; (c) fructose 1,6-bisphosphate; (d) fructose 6-phosphate 2-kinase; (e) phosphoglucoisomerase; (f) vacuolar H⁺ translocating-pyrophosphatase; (g) pyrophosphate-dependent fructose-6-phosphate phosphotransferase; (h) invertase; (i) sucrose synthase; (j) hexokinase; (k) fructokinase; (l) NDP-kinase; (m) glucose-6-phosphate 1-dehydrogenase; (n) phosphoglucomutase and (o) UDP-glucose pyrophophorylase.

The present invention also provides a substantially purified nucleic acid molecule that encodes a plant sucrose pathway enzyme or fragment thereof, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof.

The present invention also provides a substantially purified maize or soybean enzyme or fragment thereof, wherein the maize or soybean enzyme is selected from the group consisting of (a) triose phosphate isomerase; (b) fructose 1,6-bisphosphate aldolase; (c) fructose 1,6-bisphosphate; (d) fructose 6-phosphate 2-kinase; (e) phosphoglucoisomerase; (f) vacuolar H⁺ translocating-pyrophosphatase; (g) pyrophosphate-dependent fructose-6-phosphate phosphotransferase; (h) invertase; (i) sucrose synthase; (j) hexokinase; (k) fructokinase; (1) NDP-kinase; (m) glucose-6-phosphate 1-dehydrogenase; (n) phosphoglucomutase and (o) UDP-glucose pyrophophorylase.

The present invention also provides a substantially purified maize or soybean sucrose pathway protein or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

The present invention also provides a substantially purified maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate e enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified maize or soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified maize or soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified maize or soybean invertase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254.

The present invention also provides a substantially purified maize or soybean invertase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO:2254.

The present invention also provides a substantially purified maize or soybean sucrose synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified maize or soybean sucrose synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified maize or soybean hexokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified maize or soybean hexokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified maize or soybean fructokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified maize or soybean fructokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified maize or soybean NDP-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified maize or soybean NDP-kinase e enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified maize or soybean UDP-glucose pyrophophorylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a substantially purified maize or soybean UDP-glucose pyrophophorylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a purified antibody or fragment thereof which is capable of specifically binding to a maize or soybean enzyme or fragment thereof, wherein the maize or soybean enzyme or fragment thereof is encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of consisting of SEQ ID NO: 1 through SEQ ID NO: 2814.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean triose phosphate isomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 and a maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 and a maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 and a maize or soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 and a maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 and a maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 and a maize or soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 and a maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean invertase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 and a maize or soybean invertase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean sucrose synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 and a maize or soybean sucrose synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean hexokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 and a maize or soybean hexokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 and a maize or soybean fructokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean NDP-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 and a maize or soybean NDP-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 and a maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean phosphoglucomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 and a maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814 and a maize or soybean UDP-glucose pyrophophorylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; (B) a structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (a) a nucleic acid sequence which encodes for triose phosphate isomerase or fragment thereof; (b) a nucleic acid sequence which encodes for fructose 1,6-bisphosphate aldolase or fragment thereof; (c) a nucleic acid sequence which encodes for fructose 1,6-bisphosphate or fragment thereof; (d) a nucleic acid sequence which encodes for fructose 6-phosphate 2-kinase or fragment thereof; (e) a nucleic acid sequence which encodes for phosphoglucoisomerase or fragment thereof; (f) a nucleic acid sequence which encodes for vacuolar H⁺ translocating-pyrophosphatase or fragment thereof; (g) a nucleic acid sequence which encodes for pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof; (h) a nucleic acid sequence which encodes for invertase or fragment thereof; (i) a nucleic acid sequence which encodes for sucrose synthase or fragment thereof; (j) a nucleic acid sequence which encodes for hexokinase or fragment thereof; (k) a nucleic acid sequence which encodes for fructokinase or fragment thereof; (l) a nucleic acid sequence which encodes for NDP-kinase or fragment thereof; (m) a nucleic acid sequence which encodes for glucose-6-phosphate 1-dehydrogenase or fragment thereof; (n) a nucleic acid sequence which encodes for phosphoglucomutase or fragment thereof (o) a nucleic acid sequence which encodes for UDP-glucose pyrophophorylase or fragment thereof and (p) a nucleic acid sequence which is complementary to any of the nucleic acid sequences of (a) through (o); and (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule encodes a plant sucrose pathway enzyme or fragment thereof, the structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof, which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to: (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to an endogenous mRNA molecule having a nucleic acid sequence selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof, which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a method for determining a level or pattern in a plant cell of an enzyme in a plant metabolic pathway comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO: 1 through SEQ ID NO: 2814 or compliments thereof, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of an mRNA for the enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the enzyme in the plant metabolic pathway.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant sucrose pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant sucrose pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant sucrose pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or reference plant tissue with the known level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant sucrose pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or the reference plant tissue with the known level or pattern of the plant sucrose pathway enzyme.

The present invention provides a method of determining a mutation in a plant whose presence is predictive of a mutation affecting a level or pattern of a protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid, the marker nucleic acid selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having a nucleic acid sequence selected from the group of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant sucrose pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant sucrose pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method of producing a plant containing an overexpressed protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region has a nucleic acid sequence selected from group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant sucrose enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant sucrose pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof, wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant sucrose pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant sucrose pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof, wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant sucrose pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant sucrose pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either and the transcribed strand is complementary to an endogenous mRNA molecule; and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant sucrose pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to a nucleic acid molecule selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof, and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule has a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either; and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof or fragment of either and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of isolating a nucleic acid that encodes a plant sucrose pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the first nucleic acid molecule and the second nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

The present invention also provides a method of isolating a nucleic acid molecule that encodes a plant sucrose pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof or fragment of either, with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the plant sucrose pathway nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

DETAILED DESCRIPTION OF THE INVENTION

Definitions and Agents of the Present Invention

Definitions:

As used herein, a sucrose pathway enzyme is any enzyme that is associated with the synthesis or degradation of sucrose.

As used herein, a sucrose synthesis enzyme is any enzyme that is associated with the synthesis of sucrose.

As used herein, a sucrose degradation enzyme is any enzyme that is associated with the degradation of sucrose.

As used herein, triose phosphate isomerase is any enzyme that maintains at equilibrium the pool of triose phosphates, dihydroxyacetone phosphate (“DHAP”), and glyceraldehyde-3-phosphate (“GAP”) within the cytoplasm.

As used herein, fructose 1,6-bisphosphate aldolase is any enzyme that catalyzes an aldol condensation of DHAP and GAP to form fructose 1,6-bisphosphate (“F1,6BP”).

As used herein, fructose-1,6-bisphosphatase (“FBPase”) is any enzyme that catalyzes the cleavage of phosphate from the C1 carbon of fructose-1,6-bisphosphate to form fructose-6-phosphate (“F6P”).

As used herein, fructose 6-phosphate 2-kinase is any enzyme that controls the concentration of fructose 2,6-bisphosphate.

As used herein, phosphoglucoisomerase is any enzyme that maintains glucose-6-phosphate (“G6P”) and glucose-1-phosphate (“G1P”) in equilibrium with the F6P pool.

As used herein, vacuolar H⁺ translocating-pyrophosphatase is any enzyme that utilizes pyrophosphate to sustain a proton gradient formed within the vacuolar membrane.

As used herein, pyrophosphate-dependent fructose-6-phosphate phosphotransferase is any enzyme that catalyzes the reversible production of F1,6BP and Pi from F6P and PPi.

As used herein, invertase is any enzyme that irreversibly cleaves sucrose into glucose and fructose.

As used herein, sucrose synthase is any enzyme that carries out the kinetically reversible transglycosylation of sucrose and UDP into fructose and UDPG.

As used herein, hexokinase is any enzyme that can phosphorylate either glucose or fructose.

As used herein, fructokinase is any enzyme that typically has a specific affinity for fructose.

As used herein, NDP-kinase is any enzyme that can maintain UDP levels for sucrose synthase reactions, even in the case of an ATP-specific fructokinase.

As used herein, glucose-6-phosphate 1-dehydrogenase is any enzyme that allows G6P resulting from hexose kinase activity to enter the pentose phosphate pathway.

As used herein, UDP-glucose dehydrogenase is any enzyme that allows UDPG formed by sucrose synthase to be utilized directly for cellulose or callose biosynthesis.

As used herein, phosphoglucomutase is any enzyme that is ubiquitous and reversible with commitments of G6P to either F6P or G1P resulting from fluxes in metabolites further along each pathway.

Agents

(a) Nucleic Acid Molecules

Agents of the present invention include plant nucleic acid molecules and more preferably include maize and soybean nucleic acid molecules and more preferably include nucleic acid molecules of the maize genotypes B73 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), B73 x Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), DK604 (Dekalb Genetics, Dekalb, Ill. U.S.A.), H99 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), RX601 (Asgrow Seed Company, Des Moines, Iowa), Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), and soybean types Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa), C1944 (United States Department of Agriculture (USDA) Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (Tohoku University, Morioka, Japan), PI507354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.), PI227687 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.).

A subset of the nucleic acid molecules of the present invention includes nucleic acid molecules that are marker molecules. Another subset of the nucleic acid molecules of the present invention include nucleic acid molecules that encode a protein or fragment thereof. Another subset of the nucleic acid molecules of the present invention are EST molecules.

Fragment nucleic acid molecules may encode significant portion(s) of, or indeed most of, these nucleic acid molecules. Alternatively, the fragments may comprise smaller oligonucleotides (having from about 15 to about 250 nucleotide residues and more preferably, about 15 to about 30 nucleotide residues).

As used herein, an agent, be it a naturally occurring molecule or otherwise may be “substantially purified,” if desired, such that one or more molecules that is or may be present in a naturally occurring preparation containing that molecule will have been removed or will be present at a lower concentration than that at which it would normally be found.

The agents of the present invention will preferably be “biologically active” with respect to either a structural attribute, such as the capacity of a nucleic acid to hybridize to another nucleic acid molecule, or the ability of a protein to be bound by an antibody (or to compete with another molecule for such binding). Alternatively, such an attribute may be catalytic and thus involve the capacity of the agent to mediate a chemical reaction or response.

The agents of the present invention may also be recombinant. As used herein, the term recombinant means any agent (e.g. DNA, peptide etc.), that is, or results, however indirect, from human manipulation of a nucleic acid molecule.

It is understood that the agents of the present invention may be labeled with reagents that facilitate detection of the agent (e.g. fluorescent labels, Prober et al., Science 238:336-340 (1987); Albarella et al., EP 144914; chemical labels, Sheldon et al., U.S. Pat. No. 4,582,789; Albarella et al., U.S. Pat. No. 4,563,417; modified bases, Miyoshi et al., EP 119448, all of which are hereby incorporated by reference in their entirety).

It is further understood, that the present invention provides recombinant bacterial, mammalian, microbial, insect, fungal and plant cells and viral constructs comprising the agents of the present invention (See, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells).

Nucleic acid molecules or fragments thereof of the present invention are capable of specifically hybridizing to other nucleic acid molecules under certain circumstances. As used herein, two nucleic acid molecules are said to be capable of specifically hybridizing to one another if the two molecules are capable of forming an anti-parallel, double-stranded nucleic acid structure. A nucleic acid molecule is said to be the “complement” of another nucleic acid molecule if they exhibit complete complementarity. As used herein, molecules are said to exhibit “complete complementarity” when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be “minimally complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional “low-stringency” conditions. Similarly, the molecules are said to be “complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional “high-stringency” conditions. Conventional stringency conditions are described by Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989) and by Haymes et al., Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), the entirety of which is herein incorporated by reference. Departures from complete complementarity are therefore permissible, as long as such departures do not completely preclude the capacity of the molecules to form a double-stranded structure. Thus, in order for a nucleic acid molecule to serve as a primer or probe it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.

Appropriate stringency conditions which promote DNA hybridization, for example, 6.0×sodium chloride/sodium citrate (SSC) at about 45° C., followed by a wash of 2.0×SSC at 50° C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0×SSC at 50° C. to a high stringency of about 0.2×SSC at 50° C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22° C., to high stringency conditions at about 65° C. Both temperature and salt may be varied, or either the temperature or the salt concentration may be held constant while the other variable is changed.

In a preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof under moderately stringent conditions, for example at about 2.0×SSC and about 65° C.

In a particularly preferred embodiment, a nucleic acid of the present invention will include those nucleic acid molecules that specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof under high stringency conditions such as 0.2×SSC and about 65° C.

In one aspect of the present invention, the nucleic acid molecules of the present invention have one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In another aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 90% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 95% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In a more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 98% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In an even more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 99% sequence identity with one or more of the sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof.

In a further more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention exhibit 100% sequence identity with a nucleic acid molecule present within MONN01, SATMON001 through SATMON031, SATMON033, SATMON034, SATMON˜001, SATMONN01, SATMONN04 through SATMONN006, CMz029 through CMz031, CMz033, CMz035 through CMz037, CMz039 through CMz042, CMz044 through CMz045, CMz047 through CMz050, SOYMON001 through SOYMON038, Soy51 through Soy56, Soy58 through Soy62, Soy65 through Soy66, Soy 68 through Soy73 and Soy76 through Soy77, Lib9, Lib22 through Lib25, Lib35, Lib80 through Lib81, Lib 144, Lib146, Lib147, Lib190, Lib3032 through Lib3036 and Lib3099 (Monsanto Company, St. Louis, Mo. U.S.A.).

(i) Nucleic Acid Molecules Encoding Proteins or Fragments Thereof

Nucleic acid molecules of the present invention can comprise sequences that encode a sucrose pathway protein or fragment thereof. Such proteins or fragments thereof include homologues of known proteins in other organisms.

In a preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of another plant protein. In another preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of a fungal protein. In another preferred embodiment of the present invention, a maize or a soybean protein of the present invention is a homologue of mammalian protein. In another preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of a bacterial protein. In another preferred embodiment of the present invention, a soybean protein or fragment thereof of the present invention is a homologue of a maize protein. In another preferred embodiment of the present invention, a maize protein homologue or fragment thereof of the present invention is a homologue of a soybean protein.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or a soybean protein or fragment thereof where a maize or a soybean protein exhibits a BLAST probability score of greater than 1E-12, preferably a BLAST probability score of between about 1E-30 and about 1E-12, even more preferably a BLAST probability score of greater than 1E-30 with its homologue.

In another preferred embodiment of the present invention, the nucleic acid molecule encoding a maize or a soybean protein or fragment thereof exhibits a % identity with its homologue of between about 25% and about 40%, more preferably of between about 40 and about 70%, even more preferably of between about 70% and about 90% and even more preferably between about 90% and 99%. In another preferred embodiment, of the present invention, a maize or a soybean protein or fragments thereof exhibits a % identity with its homologue of 100%.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or a soybean protein or fragment thereof where a maize or a soybean protein exhibits a BLAST score of greater than 120, preferably a BLAST score of between about 1450 and about 120, even more preferably a BLAST score of greater than 1450 with its homologue.

Nucleic acid molecules of the present invention also include non-maize, non-soybean homologues. Preferred non-maize and soybean homologues are selected from the group consisting of alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm and Phaseolus.

In a preferred embodiment, nucleic acid molecules having SEQ ID NO: 1 through SEQ ID NO: 2814 or complements and fragments of either can be utilized to obtain such homologues.

The degeneracy of the genetic code, which allows different nucleic acid sequences to code for the same protein or peptide, is known in the literature. (U.S. Pat. No. 4,757,006, the entirety of which is herein incorporated by reference).

In an aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 2814 due to the degeneracy in the genetic code in that they encode the same protein but differ in nucleic acid sequence.

In another further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 2814 due to fact that the different nucleic acid sequence encodes a protein having one or more conservative amino acid residue. Examples of conservative substitutions are set forth in Table 1. It is understood that codons capable of coding for such conservative substitutions are known in the art.

TABLE 1 Original Residue Conservative Substitutions Ala Ser Arg Lys Asn Gln; His Asp Glu Cys Ser; Ala Gln Asn Glu Asp Gly Pro His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg; Gln; Glu Met Leu; Ile Phe Met; Leu; Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu

In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof due to the fact that one or more codons encoding an amino acid has been substituted for a codon that encodes a nonessential substitution of the amino acid originally encoded.

Agents of the present invention include nucleic acid molecules that encode a maize or a soybean sucrose pathway protein or fragment thereof and particularly substantially purified nucleic acid molecules selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase protein or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase protein or fragment thereof.

Non-limiting examples of such nucleic acid molecules of the present invention are nucleic acid molecules comprising: SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof that encode for a sucrose pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 or fragment thereof that encode for a triose phosphate isomerase protein or fragment thereof, SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 or fragment thereof that encode for a fructose 1,6-bisphosphate aldolase protein or fragment thereof, SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 or fragment thereof that encode for a fructose 1,6-bisphosphate protein or fragment thereof, SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 or fragment thereof that encode for a fructose 6-phosphate 2-kinase protein or fragment thereof, SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 or fragment thereof that encode for a phosphoglucoisomerase protein or fragment thereof, SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 or fragment thereof that encode for a vacuolar H⁺ translocating-pyrophosphatase protein or fragment thereof, SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 or fragment thereof that encode for a pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 or fragment thereof that encode for an invertase protein or fragment thereof, SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 or fragment thereof that encode for a sucrose synthase protein or fragment thereof, SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 or fragment thereof that encode for a hexokinase protein or fragment thereof, SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 or fragment thereof that encode for a fructokinase protein or fragment thereof, SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 or fragment thereof that encode for a NDP-kinase protein or fragment thereof, SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 or fragment thereof that encode for a glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 or fragment thereof that encode for a phosphoglucomutase protein or fragment thereof and SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814 or fragment thereof that encode for an UDP-glucose pyrophophorylase protein or fragment thereof.

A nucleic acid molecule of the present invention can also encode a homologue of a maize or a soybean triose phosphate isomerase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate aldolase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate or fragment thereof, a maize or a soybean fructose 6-phosphate 2-kinase or fragment thereof, a maize or a soybean phosphoglucoisomerase or fragment thereof, a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase or fragment thereof, a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof, a maize or a soybean invertase or fragment thereof, a maize or a soybean sucrose synthase or fragment thereof, a maize or a soybean hexokinase or fragment thereof, a maize or a soybean fructokinase or fragment thereof, a maize or a soybean NDP-kinase or fragment thereof, a maize or a soybean glucose-6-phosphate 1-dehydrogenase or fragment thereof, a maize or a soybean phosphoglucomutase or fragment thereof and a maize or a soybean UDP-glucose pyrophophorylase or fragment thereof. As used herein a homologue protein molecule or fragment thereof is a counterpart protein molecule or fragment thereof in a second species (e.g., maize triose phosphate isomerase protein is a homologue of soybean triose phosphate isomerase protein).

(ii) Nucleic Acid Molecule Markers and Probes

One aspect of the present invention concerns markers that include nucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either that can act as markers or other nucleic acid molecules of the present invention that can act as markers. Genetic markers of the present invention include “dominant” or “codominant” markers “Codominant markers” reveal the presence of two or more alleles (two per diploid individual) at a locus. “Dominant markers” reveal the presence of only a single allele per locus. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g. absence of a DNA band) is merely evidence that “some other” undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominately dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multi-allelic, codominant markers often become more informative of the genotype than dominant markers. Marker molecules can be, for example, capable of detecting polymorphisms such as single nucleotide polymorphisms (SNPs).

SNPs are single base changes in genomic DNA sequence. They occur at greater frequency and are spaced with a greater uniformly throughout a genome than other reported forms of polymorphism. The greater frequency and uniformity of SNPs means that there is greater probability that such a polymorphism will be found near or in a genetic locus of interest than would be the case for other polymorphisms. SNPs are located in protein-coding regions and noncoding regions of a genome. Some of these SNPs may result in defective or variant protein expression (e.g., as a result of mutations or defective splicing). Analysis (genotyping) of characterized SNPs can require only a plus/minus assay rather than a lengthy measurement, permitting easier automation.

SNPs can be characterized using any of a variety of methods. Such methods include the direct or indirect sequencing of the site, the use of restriction enzymes (Botstein et al., Am. J. Hum. Genet. 32:314-331 (1980), the entirety of which is herein incorporated reference; Konieczny and Ausubel, Plant J. 4:403-410 (1993), the entirety of which is herein incorporated by reference), enzymatic and chemical mismatch assays (Myers et al., Nature 313:495-498 (1985), the entirety of which is herein incorporated by reference), allele-specific PCR (Newton et al., Nucl. Acids Res. 17:2503-2516 (1989), the entirety of which is herein incorporated by reference; Wu et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:2757-2760 (1989), the entirety of which is herein incorporated by reference), ligase chain reaction (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference), single-strand conformation polymorphism analysis (Labrune et al., Am. J. Hum. Genet. 48: 1115-1120 (1991), the entirety of which is herein incorporated by reference), primer-directed nucleotide incorporation assays (Kuppuswami et al., Proc. Natl. Acad. Sci. USA 88:1143-1147 (1991), the entirety of which is herein incorporated by reference), dideoxy fingerprinting (Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference), solid-phase ELISA-based oligonucleotide ligation assays (Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the entirety of which is herein incorporated by reference), oligonucleotide fluorescence-quenching assays (Livak et al., PCR Methods Appl. 4:357-362 (1995), the entirety of which is herein incorporated by reference), 5′-nuclease allele-specific hybridization TaqMan assay (Livak et al., Nature Genet. 9:341-342 (1995), the entirety of which is herein incorporated by reference), template-directed dye-terminator incorporation (TDI) assay (Chen and Kwok, Nucl. Acids Res. 25:347-353 (1997), the entirety of which is herein incorporated by reference), allele-specific molecular beacon assay (Tyagi et al., Nature Biotech. 16: 49-53 (1998), the entirety of which is herein incorporated by reference), PinPoint assay (Haff and Smirnov, Genome Res. 7: 378-388 (1997), the entirety of which is herein incorporated by reference) and dCAPS analysis (Neff et al., Plant J. 14:387-392 (1998), the entirety of which is herein incorporated by reference).

Additional markers, such as AFLP markers, RFLP markers and RAPD markers, can be utilized (Walton, Seed World 22-29 (July, 1993), the entirety of which is herein incorporated by reference; Burow and Blake, Molecular Dissection of Complex Traits, 13-29, Paterson (ed.), CRC Press, New York (1988), the entirety of which is herein incorporated by reference). DNA markers can be developed from nucleic acid molecules using restriction endonucleases, the PCR and/or DNA sequence information. RFLP markers result from single base changes or insertions/deletions. These codominant markers are highly abundant in plant genomes, have a medium level of polymorphism and are developed by a combination of restriction endonuclease digestion and Southern blotting hybridization. CAPS are similarly developed from restriction nuclease digestion but only of specific PCR products. These markers are also codominant, have a medium level of polymorphism and are highly abundant in the genome. The CAPS result from single base changes and insertions/deletions.

Another marker type, RAPDs, are developed from DNA amplification with random primers and result from single base changes and insertions/deletions in plant genomes. They are dominant markers with a medium level of polymorphisms and are highly abundant. AFLP markers require using the PCR on a subset of restriction fragments from extended adapter primers. These markers are both dominant and codominant are highly abundant in genomes and exhibit a medium level of polymorphism.

SSRs require DNA sequence information. These codominant markers result from repeat length changes, are highly polymorphic and do not exhibit as high a degree of abundance in the genome as CAPS, AFLPs and RAPDs SNPs also require DNA sequence information. These codominant markers result from single base substitutions. They are highly abundant and exhibit a medium of polymorphism (Rafalski et al., In: Nonmammalian Genomic Analysis, Birren and Lai (ed.), Academic Press, San Diego, Calif., pp. 75-134 (1996), the entirety of which is herein incorporated by reference). It is understood that a nucleic acid molecule of the present invention may be used as a marker.

A PCR probe is a nucleic acid molecule capable of initiating a polymerase activity while in a double-stranded structure to with another nucleic acid. Various methods for determining the structure of PCR probes and PCR techniques exist in the art. Computer generated searches using programs such as Primer3 available on the worldwide web at genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline available on the worldwide web at genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole et al., BioTechniques 25:112-123 (1998) the entirety of which is herein incorporated by reference), for example, can be used to identify potential PCR primers.

It is understood that a fragment of one or more of the nucleic acid molecules of the present invention may be a probe and specifically a PCR probe.

(b) Protein and Peptide Molecules

A class of agents comprises one or more of the protein or fragments thereof or peptide molecules encoded by SEQ ID NO: 1 through SEQ ID NO: 2814 or one or more of the protein or fragment thereof and peptide molecules encoded by other nucleic acid agents of the present invention. As used herein, the term “protein molecule” or “peptide molecule” includes any molecule that comprises five or more amino acids. It is well known in the art that proteins may undergo modification, including post-translational modifications, such as, but not limited to, disulfide bond formation, glycosylation, phosphorylation, or oligomerization. Thus, as used herein, the term “protein molecule” or “peptide molecule” includes any protein molecule that is modified by any biological or non-biological process. The terms “amino acid” and “amino acids” refer to all naturally occurring L-amino acids. This definition is meant to include norleucine, ornithine, homocysteine and homoserine.

Non-limiting examples of the protein or fragment thereof of the present invention include a maize or a soybean sucrose pathway protein or fragment thereof; a maize or a soybean triose phosphate isomerase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate aldolase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate or fragment thereof, a maize or a soybean fructose 6-phosphate 2-kinase or fragment thereof, a maize or a soybean phosphoglucoisomerase or fragment thereof, a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase or fragment thereof, a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof, a maize or a soybean invertase or fragment thereof, a maize or a soybean sucrose synthase or fragment thereof, a maize or a soybean hexokinase or fragment thereof, a maize or a soybean fructokinase or fragment thereof, a maize or a soybean NDP-kinase or fragment thereof, a maize or a soybean glucose-6-phosphate 1-dehydrogenase or fragment thereof, a maize or a soybean phosphoglucomutase or fragment thereof and a maize or a soybean UDP-glucose pyrophophorylase or fragment thereof.

Non-limiting examples of the protein or fragment molecules of the present invention are a sucrose pathway protein or fragment thereof encoded by: SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof that encode for a sucrose pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 or fragment thereof that encode for a triose phosphate isomerase protein or fragment thereof, SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 or fragment thereof that encode for a fructose 1,6-bisphosphate aldolase protein or fragment thereof, SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 or fragment thereof that encode for a fructose 1,6-bisphosphate protein or fragment thereof, SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 or fragment thereof that encode for a fructose 6-phosphate 2-kinase protein or fragment thereof, SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 or fragment thereof that encode for a phosphoglucoisomerase protein or fragment thereof, SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 or fragment thereof that encode for a vacuolar H⁺ translocating-pyrophosphatase protein or fragment thereof, SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 or fragment thereof that encode for a pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 or fragment thereof that encode for an invertase protein or fragment thereof, SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 or fragment thereof that encode for a sucrose synthase protein or fragment thereof, SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 or fragment thereof that encode for a hexokinase protein or fragment thereof, SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 or fragment thereof that encode for a fructokinase protein or fragment thereof, SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 or fragment thereof that encode for a NDP-kinase protein or fragment thereof, SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 or fragment thereof that encode for a glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 or fragment thereof that encode for a phosphoglucomutase protein or fragment thereof and SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814 or fragment thereof that encode for an UDP-glucose pyrophophorylase protein or fragment thereof.

One or more of the protein or fragment of peptide molecules may be produced via chemical synthesis, or more preferably, by expressing in a suitable bacterial or eucaryotic host. Suitable methods for expression are described by Sambrook et al., (In: Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)), or similar texts. For example, the protein may be expressed in, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells.

A “protein fragment” is a peptide or polypeptide molecule whose amino acid sequence comprises a subset of the amino acid sequence of that protein. A protein or fragment thereof that comprises one or more additional peptide regions not derived from that protein is a “fusion” protein. Such molecules may be derivatized to contain carbohydrate or other moieties (such as keyhole limpet hemocyanin, etc.). Fusion protein or peptide molecules of the present invention are preferably produced via recombinant means.

Another class of agents comprise protein or peptide molecules or fragments or fusions thereof encoded by SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof in which conservative, non-essential or non-relevant amino acid residues have been added, replaced or deleted. Computerized means for designing modifications in protein structure are known in the art (Dahiyat and Mayo, Science 278:82-87 (1997), the entirety of which is herein incorporated by reference).

The protein molecules of the present invention include plant homologue proteins. An example of such a homologue is a homologue protein of a non-maize or non-soybean plant species, that include but not limited to alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus etc. Particularly preferred non-maize or non-soybean for use for the isolation of homologs would include, Arabidopsis, barley, cotton, oat, oilseed rape, rice, canola, ornamentals, sugarcane, sugarbeet, tomato, potato, wheat and turf grasses. Such a homologue can be obtained by any of a variety of methods. Most preferably, as indicated above, one or more of the disclosed sequences (SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof) will be used to define a pair of primers that may be used to isolate the homologue-encoding nucleic acid molecules from any desired species. Such molecules can be expressed to yield homologues by recombinant means.

(c) Antibodies

One aspect of the present invention concerns antibodies, single-chain antigen binding molecules, or other proteins that specifically bind to one or more of the protein or peptide molecules of the present invention and their homologues, fusions or fragments. Such antibodies may be used to quantitatively or qualitatively detect the protein or peptide molecules of the present invention. As used herein, an antibody or peptide is said to “specifically bind” to a protein or peptide molecule of the present invention if such binding is not competitively inhibited by the presence of non-related molecules.

Nucleic acid molecules that encode all or part of the protein of the present invention can be expressed, via recombinant means, to yield protein or peptides that can in turn be used to elicit antibodies that are capable of binding the expressed protein or peptide. Such antibodies may be used in immunoassays for that protein. Such protein-encoding molecules, or their fragments may be a “fusion” molecule (i.e., a part of a larger nucleic acid molecule) such that, upon expression, a fusion protein is produced. It is understood that any of the nucleic acid molecules of the present invention may be expressed, via recombinant means, to yield proteins or peptides encoded by these nucleic acid molecules.

The antibodies that specifically bind proteins and protein fragments of the present invention may be polyclonal or monoclonal and may comprise intact immunoglobulins, or antigen binding portions of immunoglobulins fragments (such as (F(ab′), F(ab′)₂), or single-chain immunoglobulins producible, for example, via recombinant means. It is understood that practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of antibodies (see, for example, Harlow and Lane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988), the entirety of which is herein incorporated by reference).

Murine monoclonal antibodies are particularly preferred. BALB/c mice are preferred for this purpose, however, equivalent strains may also be used. The animals are preferably immunized with approximately 25 μg of purified protein (or fragment thereof) that has been emulsified in a suitable adjuvant (such as TiterMax adjuvant (Vaxcel, Norcross, Ga.)). Immunization is preferably conducted at two intramuscular sites, one intraperitoneal site and one subcutaneous site at the base of the tail. An additional i.v. injection of approximately 25 μg of antigen is preferably given in normal saline three weeks later. After approximately 11 days following the second injection, the mice may be bled and the blood screened for the presence of anti-protein or peptide antibodies. Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) is employed for this purpose.

More preferably, the mouse having the highest antibody titer is given a third i.v. injection of approximately 25 μg of the same protein or fragment. The splenic leukocytes from this animal may be recovered 3 days later and then permitted to fuse, most preferably, using polyethylene glycol, with cells of a suitable myeloma cell line (such as, for example, the P3×63Ag8.653 myeloma cell line). Hybridoma cells are selected by culturing the cells under “HAT” (hypoxanthine-aminopterin-thymine) selection for about one week. The resulting clones may then be screened for their capacity to produce monoclonal antibodies (“mAbs”), preferably by direct ELISA.

In one embodiment, anti-protein or peptide monoclonal antibodies are isolated using a fusion of a protein or peptide of the present invention, or conjugate of a protein or peptide of the present invention, as immunogens. Thus, for example, a group of mice can be immunized using a fusion protein emulsified in Freund's complete adjuvant (e.g. approximately 50 μg of antigen per immunization). At three week intervals, an identical amount of antigen is emulsified in Freund's incomplete adjuvant and used to immunize the animals. Ten days following the third immunization, serum samples are taken and evaluated for the presence of antibody. If antibody titers are too low, a fourth booster can be employed. Polysera capable of binding the protein or peptide can also be obtained using this method.

In a preferred procedure for obtaining monoclonal antibodies, the spleens of the above-described immunized mice are removed, disrupted and immune splenocytes are isolated over a ficoll gradient. The isolated splenocytes are fused, using polyethylene glycol with BALB/c-derived HGPRT (hypoxanthine guanine phosphoribosyl transferase) deficient P3×63×Ag8.653 plasmacytoma cells. The fused cells are plated into 96 well microtiter plates and screened for hybridoma fusion cells by their capacity to grow in culture medium supplemented with hypothanthine, aminopterin and thymidine for approximately 2-3 weeks.

Hybridoma cells that arise from such incubation are preferably screened for their capacity to produce an immunoglobulin that binds to a protein of interest. An indirect ELISA may be used for this purpose. In brief, the supernatants of hybridomas are incubated in microtiter wells that contain immobilized protein. After washing, the titer of bound immunoglobulin can be determined using, for example, a goat anti-mouse antibody conjugated to horseradish peroxidase. After additional washing, the amount of immobilized enzyme is determined (for example through the use of a chromogenic substrate). Such screening is performed as quickly as possible after the identification of the hybridoma in order to ensure that a desired clone is not overgrown by non-secreting neighbor cells. Desirably, the fusion plates are screened several times since the rates of hybridoma growth vary. In a preferred sub-embodiment, a different antigenic form may be used to screen the hybridoma. Thus, for example, the splenocytes may be immunized with one immunogen, but the resulting hybridomas can be screened using a different immunogen. It is understood that any of the protein or peptide molecules of the present invention may be used to raise antibodies.

As discussed below, such antibody molecules or their fragments may be used for diagnostic purposes. Where the antibodies are intended for diagnostic purposes, it may be desirable to derivatize them, for example with a ligand group (such as biotin) or a detectable marker group (such as a fluorescent group, a radioisotope or an enzyme).

The ability to produce antibodies that bind the protein or peptide molecules of the present invention permits the identification of mimetic compounds of those molecules. A “mimetic compound” is a compound that is not that compound, or a fragment of that compound, but which nonetheless exhibits an ability to specifically bind to antibodies directed against that compound.

It is understood that any of the agents of the present invention can be substantially purified and/or be biologically active and/or recombinant.

Uses of the Agents of the Invention

Nucleic acid molecules and fragments thereof of the present invention may be employed to obtain other nucleic acid molecules from the same species (e.g., ESTs or fragment thereof from maize may be utilized to obtain other nucleic acid molecules from maize). Such nucleic acid molecules include the nucleic acid molecules that encode the complete coding sequence of a protein and promoters and flanking sequences of such molecules. In addition, such nucleic acid molecules include nucleic acid molecules that encode for other isozymes or gene family members. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from maize or soybean. Methods for forming such libraries are well known in the art.

Nucleic acid molecules and fragments thereof of the present invention may also be employed to obtain nucleic acid homologues. Such homologues include the nucleic acid molecule of other plants or other organisms (e.g., alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus, etc.) including the nucleic acid molecules that encode, in whole or in part, protein homologues of other plant species or other organisms, sequences of genetic elements such as promoters and transcriptional regulatory elements. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from such plant species. Methods for forming such libraries are well known in the art. Such homologue molecules may differ in their nucleotide sequences from those found in one or more of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof because complete complementarity is not needed for stable hybridization. The nucleic acid molecules of the present invention therefore also include molecules that, although capable of specifically hybridizing with the nucleic acid molecules, may lack “complete complementarity.”

Any of a variety of methods may be used to obtain one or more of the above-described nucleic acid molecules (Zamechik et al., Proc. Natl. Acad. Sci. (U.S.A.) 83:4143-4146 (1986), the entirety of which is herein incorporated by reference; Goodchild et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:5507-5511 (1988), the entirety of which is herein incorporated by reference; Wickstrom et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1028-1032 (1988), the entirety of which is herein incorporated by reference; Holt et al., Molec. Cell. Biol. 8:963-973 (1988), the entirety of which is herein incorporated by reference; Gerwirtz et al., Science 242:1303-1306 (1988), the entirety of which is herein incorporated by reference; Anfossi et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:3379-3383 (1989), the entirety of which is herein incorporated by reference; Becker et al., EMBO J. 8:3685-3691 (1989); the entirety of which is herein incorporated by reference). Automated nucleic acid synthesizers may be employed for this purpose. In lieu of such synthesis, the disclosed nucleic acid molecules may be used to define a pair of primers that can be used with the polymerase chain reaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; Mullis, European Patent 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194, all of which are herein incorporated by reference in their entirety) to amplify and obtain any desired nucleic acid molecule or fragment.

Promoter sequence(s) and other genetic elements, including but not limited to transcriptional regulatory flanking sequences, associated with one or more of the disclosed nucleic acid sequences can also be obtained using the disclosed nucleic acid sequence provided herein. In one embodiment, such sequences are obtained by incubating EST nucleic acid molecules or preferably fragments thereof with members of genomic libraries (e.g. maize and soybean) and recovering clones that hybridize to the EST nucleic acid molecule or fragment thereof. In a second embodiment, methods of “chromosome walking,” or inverse PCR may be used to obtain such sequences (Frohman et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048 (1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et al., Method Mol. Biol. 67:287-294 (1997); Benkel et al., Genet. Anal. 13:123-127 (1996); Hartl et al., Methods Mol. Biol. 58:293-301 (1996), all of which are herein incorporated by reference in their entirety).

The nucleic acid molecules of the present invention may be used to isolate promoters of cell enhanced, cell specific, tissue enhanced, tissue specific, developmentally or environmentally regulated expression profiles. Isolation and functional analysis of the 5′ flanking promoter sequences of these genes from genomic libraries, for example, using genomic screening methods and PCR techniques would result in the isolation of useful promoters and transcriptional regulatory elements. These methods are known to those of skill in the art and have been described (See, for example, Birren et al., Genome Analysis: Analyzing DNA, 1, (1997), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference). Promoters obtained utilizing the nucleic acid molecules of the present invention could also be modified to affect their control characteristics. Examples of such modifications would include but are not limited to enhanced sequences as reported in Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants. Such genetic elements could be used to enhance gene expression of new and existing traits for crop improvements.

In one sub-aspect, such an analysis is conducted by determining the presence and/or identity of polymorphism(s) by one or more of the nucleic acid molecules of the present invention and more preferably one or more of the EST nucleic acid molecule or fragment thereof which are associated with a phenotype, or a predisposition to that phenotype.

Any of a variety of molecules can be used to identify such polymorphism(s). In one embodiment, one or more of the EST nucleic acid molecules (or a sub-fragment thereof) may be employed as a marker nucleic acid molecule to identify such polymorphism(s). Alternatively, such polymorphisms can be detected through the use of a marker nucleic acid molecule or a marker protein that is genetically linked to (i.e., a polynucleotide that co-segregates with) such polymorphism(s).

In an alternative embodiment, such polymorphisms can be detected through the use of a marker nucleic acid molecule that is physically linked to such polymorphism(s). For this purpose, marker nucleic acid molecules comprising a nucleotide sequence of a polynucleotide located within 1 mb of the polymorphism(s) and more preferably within 100 kb of the polymorphism(s) and most preferably within 10 kb of the polymorphism(s) can be employed.

The genomes of animals and plants naturally undergo spontaneous mutation in the course of their continuing evolution (Gusella, Ann. Rev. Biochem. 55:831-854 (1986)). A “polymorphism” is a variation or difference in the sequence of the gene or its flanking regions that arises in some of the members of a species. The variant sequence and the “original” sequence co-exist in the species' population. In some instances, such co-existence is in stable or quasi-stable equilibrium.

A polymorphism is thus said to be “allelic,” in that, due to the existence of the polymorphism, some members of a species may have the original sequence (i.e., the original “allele”) whereas other members may have the variant sequence (i.e., the variant “allele”). In the simplest case, only one variant sequence may exist and the polymorphism is thus said to be di-allelic. In other cases, the species' population may contain multiple alleles and the polymorphism is termed tri-allelic, etc. A single gene may have multiple different unrelated polymorphisms. For example, it may have a di-allelic polymorphism at one site and a multi-allelic polymorphism at another site.

The variation that defines the polymorphism may range from a single nucleotide variation to the insertion or deletion of extended regions within a gene. In some cases, the DNA sequence variations are in regions of the genome that are characterized by short tandem repeats (STRs) that include tandem di- or tri-nucleotide repeated motifs of nucleotides. Polymorphisms characterized by such tandem repeats are referred to as “variable number tandem repeat” (“VNTR”) polymorphisms. VNTRs have been used in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour et al., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol. 39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003; Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat. No. 5,175,082; Jeffreys et al., Amer. J. Hum. Genet. 39:11-24 (1986); Jeffreys et al., Nature 316:76-79 (1985); Gray et al., Proc. R. Acad. Soc. Lond. 243:241-253 (1991); Moore et al., Genomics 10:654-660 (1991); Jeffreys et al., Anim. Genet. 18:1-15 (1987); Hillel et al., Anim. Genet. 20:145-155 (1989); Hillel et al., Genet. 124:783-789 (1990), all of which are herein incorporated by reference in their entirety).

The detection of polymorphic sites in a sample of DNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis or other means.

The most preferred method of achieving such amplification employs the polymerase chain reaction (“PCR”) (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent Appln. 50,424; European Patent Appln. 84,796; European Patent Application 258,017; European Patent Appln. 237,362; Mullis, European Patent Appln. 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

In lieu of PCR, alternative methods, such as the “Ligase Chain Reaction” (“LCR”) may be used (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference). LCR uses two pairs of oligonucleotide probes to exponentially amplify a specific target. The sequences of each pair of oligonucleotides is selected to permit the pair to hybridize to abutting sequences of the same strand of the target. Such hybridization forms a substrate for a template-dependent ligase. As with PCR, the resulting products thus serve as a template in subsequent cycles and an exponential amplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal and distal sequences of the same strand of a polymorphic site. In one embodiment, either oligonucleotide will be designed to include the actual polymorphic site of the polymorphism. In such an embodiment, the reaction conditions are selected such that the oligonucleotides can be ligated together only if the target molecule either contains or lacks the specific nucleotide that is complementary to the polymorphic site present on the oligonucleotide. Alternatively, the oligonucleotides may be selected such that they do not include the polymorphic site (see, Segev, PCT Application WO 90/01069, the entirety of which is herein incorporated by reference).

The “Oligonucleotide Ligation Assay” (“OLA”) may alternatively be employed (Landegren et al., Science 241:1077-1080 (1988), the entirety of which is herein incorporated by reference). The OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target. OLA, like LCR, is particularly suited for the detection of point mutations. Unlike LCR, however, OLA results in “linear” rather than exponential amplification of the target sequence.

Nickerson et al., have described a nucleic acid detection assay that combines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8923-8927 (1990), the entirety of which is herein incorporated by reference). In this method, PCR is used to achieve the exponential amplification of target DNA, which is then detected using OLA. In addition to requiring multiple and separate, processing steps, one problem associated with such combinations is that they inherit all of the problems associated with PCR and OLA.

Schemes based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting “di-oligonucleotide”, thereby amplifying the di-oligonucleotide, are also known (Wu et al., Genomics 4:560-569 (1989), the entirety of which is herein incorporated by reference) and may be readily adapted to the purposes of the present invention.

Other known nucleic acid amplification procedures, such as allele-specific oligomers, branched DNA technology, transcription-based amplification systems, or isothermal amplification methods may also be used to amplify and analyze such polymorphisms (Malek et al., U.S. Pat. No. 5,130,238; Davey et al., European Patent Application 329,822; Schuster et al., U.S. Pat. No. 5,169,766; Miller et al., PCT Patent Application WO 89/06700; Kwoh et al, Proc. Natl. Acad. Sci. (U.S.A.) 86:1173-1177 (1989); Gingeras et al., PCT Patent Application WO 88/10315; Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:392-396 (1992), all of which are herein incorporated by reference in their entirety).

The identification of a polymorphism can be determined in a variety of ways. By correlating the presence or absence of it in a plant with the presence or absence of a phenotype, it is possible to predict the phenotype of that plant. If a polymorphism creates or destroys a restriction endonuclease cleavage site, or if it results in the loss or insertion of DNA (e.g., a VNTR polymorphism), it will alter the size or profile of the DNA fragments that are generated by digestion with that restriction endonuclease. As such, individuals that possess a variant sequence can be distinguished from those having the original sequence by restriction fragment analysis. Polymorphisms that can be identified in this manner are termed “restriction fragment length polymorphisms” (“RFLPs”). RFLPs have been widely used in human and plant genetic analyses (Glassberg, UK Patent Application 2135774; Skolnick et al., Cytogen. Cell Genet. 32:58-67 (1982); Botstein et al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer et al., (PCT Application WO90/13668); Uhlen, PCT Application WO90/11369).

Polymorphisms can also be identified by Single Strand Conformation Polymorphism (SSCP) analysis. SSCP is a method capable of identifying most sequence variations in a single strand of DNA, typically between 150 and 250 nucleotides in length (Elles, Methods in Molecular Medicine: Molecular Diagnosis of Genetic Diseases, Humana Press (1996), the entirety of which is herein incorporated by reference); Orita et al., Genomics 5:874-879 (1989), the entirety of which is herein incorporated by reference). Under denaturing conditions a single strand of DNA will adopt a conformation that is uniquely dependent on its sequence conformation. This conformation usually will be different, even if only a single base is changed. Most conformations have been reported to alter the physical configuration or size sufficiently to be detectable by electrophoresis. A number of protocols have been described for SSCP including, but not limited to, Lee et al., Anal. Biochem. 205:289-293 (1992), the entirety of which is herein incorporated by reference; Suzuki et al., Anal. Biochem. 192:82-84 (1991), the entirety of which is herein incorporated by reference; Lo et al., Nucleic Acids Research 20:1005-1009 (1992), the entirety of which is herein incorporated by reference; Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference. It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by SSCP analysis.

Polymorphisms may also be found using a DNA fingerprinting technique called amplified fragment length polymorphism (AFLP), which is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA to profile that DNA (Vos et al., Nucleic Acids Res. 23:4407-4414 (1995), the entirety of which is herein incorporated by reference). This method allows for the specific co-amplification of high numbers of restriction fragments, which can be visualized by PCR without knowledge of the nucleic acid sequence.

AFLP employs basically three steps. Initially, a sample of genomic DNA is cut with restriction enzymes and oligonucleotide adapters are ligated to the restriction fragments of the DNA. The restriction fragments are then amplified using PCR by using the adapter and restriction sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotide flanking the restriction sites. These amplified fragments are then visualized on a denaturing polyacrylamide gel.

AFLP analysis has been performed on Salix (Beismann et al., Mol. Ecol. 6:989-993 (1997), the entirety of which is herein incorporated by reference), Acinetobacter (Janssen et al., Int. J. Syst. Bacteriol. 47:1179-1187 (1997), the entirety of which is herein incorporated by reference), Aeromonas popoffi (Huys et al., Int. J. Syst. Bacteriol. 47:1165-1171 (1997), the entirety of which is herein incorporated by reference), rice (McCouch et al., Plant Mol. Biol. 35:89-99 (1997), the entirety of which is herein incorporated by reference; Nandi et al., Mol. Gen. Genet. 255:1-8 (1997), the entirety of which is herein incorporated by reference; Cho et al., Genome 39:373-378 (1996), the entirety of which is herein incorporated by reference), barley (Hordeum vulgare)(Simons et al., Genomics 44:61-70 (1997), the entirety of which is herein incorporated by reference; Waugh et al, Mol. Gen. Genet. 255:311-321 (1997), the entirety of which is herein incorporated by reference; Qi et al., Mol. Gen Genet. 254:330-336 (1997), the entirety of which is herein incorporated by reference; Becker et al., Mol. Gen. Genet. 249:65-73 (1995), the entirety of which is herein incorporated by reference), potato (Van der Voort et al., Mol. Gen. Genet. 255:438-447 (1997), the entirety of which is herein incorporated by reference; Meksem et al., Mol. Gen. Genet. 249:74-81 (1995), the entirety of which is herein incorporated by reference), Phytophthora infestans (Van der Lee et al., Fungal Genet. Biol. 21:278-291 (1997), the entirety of which is herein incorporated by reference), Bacillus anthracis (Keim et al., J. Bacteriol. 179:818-824 (1997), the entirety of which is herein incorporated by reference), Astragalus cremnophylax (Travis et al., Mol. Ecol. 5:735-745 (1996), the entirety of which is herein incorporated by reference), Arabidopsis (Cnops et al., Mol. Gen. Genet. 253:32-41 (1996), the entirety of which is herein incorporated by reference), Escherichia coli (Lin et al., Nucleic Acids Res. 24:3649-3650 (1996), the entirety of which is herein incorporated by reference), Aeromonas (Huys et al., Int. J. Syst. Bacteriol. 46:572-580 (1996), the entirety of which is herein incorporated by reference), nematode (Folkertsma et al., Mol. Plant Microbe Interact. 9:47-54 (1996), the entirety of which is herein incorporated by reference), tomato (Thomas et al., Plant J. 8:785-794 (1995), the entirety of which is herein incorporated by reference) and human (Latorra et al., PCR Methods Appl. 3:351-358 (1994), the entirety of which is herein incorporated by reference). AFLP analysis has also been used for fingerprinting mRNA (Money et al., Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which is herein incorporated by reference; Bachem et al., Plant J. 9:745-753 (1996), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by AFLP analysis or for fingerprinting RNA.

Polymorphisms may also be found using random amplified polymorphic DNA (RAPD) (Williams et al., Nucl. Acids Res. 18:6531-6535 (1990), the entirety of which is herein incorporated by reference) and cleaveable amplified polymorphic sequences (CAPS) (Lyamichev et al, Science 260:778-783 (1993), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention, may be utilized as markers or probes to detect polymorphisms by RAPD or CAPS analysis.

Through genetic mapping, a fine scale linkage map can be developed using DNA markers and, then, a genomic DNA library of large-sized fragments can be screened with molecular markers linked to the desired trait. Molecular markers are advantageous for agronomic traits that are otherwise difficult to tag, such as resistance to pathogens, insects and nematodes, tolerance to abiotic stress, quality parameters and quantitative traits such as high yield potential.

The essential requirements for marker-assisted selection in a plant breeding program are: (1) the marker(s) should co-segregate or be closely linked with the desired trait; (2) an efficient means of screening large populations for the molecular marker(s) should be available; and (3) the screening technique should have high reproducibility across laboratories and preferably be economical to use and be user-friendly.

The genetic linkage of marker molecules can be established by a gene mapping model such as, without limitation, the flanking marker model reported by Lander and Botstein, Genetics 121:185-199 (1989) and the interval mapping, based on maximum likelihood methods described by Lander and Botstein, Genetics 121:185-199 (1989) and implemented in the software package MAPMAKER/QTL (Lincoln and Lander, Mapping Genes Controlling Quantitative Traits Using MAPMAKER/QTL, Whitehead Institute for Biomedical Research, Massachusetts, (1990). Additional software includes Qgene, Version 2.23 (1996), Department of Plant Breeding and Biometry, 266 Emerson Hall, Cornell University, Ithaca, N.Y., the manual of which is herein incorporated by reference in its entirety). Use of Qgene software is a particularly preferred approach.

A maximum likelihood estimate (MLE) for the presence of a marker is calculated, together with an MLE assuming no QTL effect, to avoid false positives. A log₁₀ of an odds ratio (LOD) is then calculated as: LOD=log₁₀ (MLE for the presence of a QTL/MLE given no linked QTL).

The LOD score essentially indicates how much more likely the data are to have arisen assuming the presence of a QTL than in its absence. The LOD threshold value for avoiding a false positive with a given confidence, say 95%, depends on the number of markers and the length of the genome. Graphs indicating LOD thresholds are set forth in Lander and Botstein, Genetics 121:185-199 (1989) the entirety of which is herein incorporated by reference and further described by Arús and Moreno-González, Plant Breeding, Hayward et al., (eds.) Chapman & Hall, London, pp. 314-331 (1993), the entirety of which is herein incorporated by reference.

Additional models can be used. Many modifications and alternative approaches to interval mapping have been reported, including the use non-parametric methods (Kruglyak and Lander, Genetics 139:1421-1428 (1995), the entirety of which is herein incorporated by reference). Multiple regression methods or models can be also be used, in which the trait is regressed on a large number of markers (Jansen, Biometrics in Plant Breeding, van Oijen and Jansen (eds.), Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 116-124 (1994); Weber and Wricke, Advances in Plant Breeding, Blackwell, Berlin, 16 (1994), both of which is herein incorporated by reference in their entirety). Procedures combining interval mapping with regression analysis, whereby the phenotype is regressed onto a single putative QTL at a given marker interval and at the same time onto a number of markers that serve as ‘cofactors,’ have been reported by Jansen and Stam, Genetics 136:1447-1455 (1994), the entirety of which is herein incorporated by reference and Zeng, Genetics 136:1457-1468 (1994) the entirety of which is herein incorporated by reference. Generally, the use of cofactors reduces the bias and sampling error of the estimated QTL positions (Utz and Melchinger, Biometrics in Plant Breeding, van Oijen and Jansen (eds.) Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp.195-204 (1994), the entirety of which is herein incorporated by reference, thereby improving the precision and efficiency of QTL mapping (Zeng, Genetics 136:1457-1468 (1994)). These models can be extended to multi-environment experiments to analyze genotype-environment interactions (Jansen et al., Theo. Appl. Genet. 91:33-37 (1995), the entirety of which is herein incorporated by reference).

Selection of an appropriate mapping populations is important to map construction. The choice of appropriate mapping population depends on the type of marker systems employed (Tanksley et al., Molecular mapping plant chromosomes. Chromosome structure and function: Impact of new concepts, Gustafson and Appels (eds.), Plenum Press, New York, pp 157-173 (1988), the entirety of which is herein incorporated by reference). Consideration must be given to the source of parents (adapted vs. exotic) used in the mapping population. Chromosome pairing and recombination rates can be severely disturbed (suppressed) in wide crosses (adapted×exotic) and generally yield greatly reduced linkage distances. Wide crosses will usually provide segregating populations with a relatively large array of polymorphisms when compared to progeny in a narrow cross (adapted×adapted).

An F₂ population is the first generation of selfing after the hybrid seed is produced. Usually a single F₁ plant is selfed to generate a population segregating for all the genes in Mendelian (1:2:1) fashion. Maximum genetic information is obtained from a completely classified F₂ population using a codominant marker system (Mather, Measurement of Linkage in Heredity, Methuen and Co., (1938), the entirety of which is herein incorporated by reference). In the case of dominant markers, progeny tests (e.g. F₃, BCF₂) are required to identify the heterozygotes, thus making it equivalent to a completely classified F₂ population. However, this procedure is often prohibitive because of the cost and time involved in progeny testing. Progeny testing of F₂ individuals is often used in map construction where phenotypes do not consistently reflect genotype (e.g. disease resistance) or where trait expression is controlled by a QTL. Segregation data from progeny test populations (e.g. F₃ or BCF₂) can be used in map construction. Marker-assisted selection can then be applied to cross progeny based on marker-trait map associations (F₂, F₃), where linkage groups have not been completely disassociated by recombination events (i.e., maximum disequillibrium).

Recombinant inbred lines (RIL) (genetically related lines; usually >F₅, developed from continuously selfing F₂ lines towards homozygosity) can be used as a mapping population. Information obtained from dominant markers can be maximized by using RIL because all loci are homozygous or nearly so. Under conditions of tight linkage (i.e., about <10% recombination), dominant and co-dominant markers evaluated in RIL populations provide more information per individual than either marker type in backcross populations (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992), the entirety of which is herein incorporated by reference). However, as the distance between markers becomes larger (i.e., loci become more independent), the information in RIL populations decreases dramatically when compared to codominant markers.

Backcross populations (e.g., generated from a cross between a successful variety (recurrent parent) and another variety (donor parent) carrying a trait not present in the former) can be utilized as a mapping population. A series of backcrosses to the recurrent parent can be made to recover most of its desirable traits. Thus a population is created consisting of individuals nearly like the recurrent parent but each individual carries varying amounts or mosaic of genomic regions from the donor parent. Backcross populations can be useful for mapping dominant markers if all loci in the recurrent parent are homozygous and the donor and recurrent parent have contrasting polymorphic marker alleles (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)). Information obtained from backcross populations using either codominant or dominant markers is less than that obtained from F₂ populations because one, rather than two, recombinant gametes are sampled per plant. Backcross populations, however, are more informative (at low marker saturation) when compared to RILs as the distance between linked loci increases in RIL populations (i.e. about 15% recombination). Increased recombination can be beneficial for resolution of tight linkages, but may be undesirable in the construction of maps with low marker saturation.

Near-isogenic lines (NIL) created by many backcrosses to produce an array of individuals that are nearly identical in genetic composition except for the trait or genomic region under interrogation can be used as a mapping population. In mapping with NILs, only a portion of the polymorphic loci are expected to map to a selected region.

Bulk segregant analysis (BSA) is a method developed for the rapid identification of linkage between markers and traits of interest (Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9828-9832 (1991), the entirety of which is herein incorporated by reference). In BSA, two bulked DNA samples are drawn from a segregating population originating from a single cross. These bulks contain individuals that are identical for a particular trait (resistant or susceptible to particular disease) or genomic region but arbitrary at unlinked regions (i.e. heterozygous). Regions unlinked to the target region will not differ between the bulked samples of many individuals in BSA.

It is understood that one or more of the nucleic acid molecules of the present invention may be used as molecular markers. It is also understood that one or more of the protein molecules of the present invention may be used as molecular markers.

In accordance with this aspect of the present invention, a sample nucleic acid is obtained from plants cells or tissues. Any source of nucleic acid may be used. Preferably, the nucleic acid is genomic DNA. The nucleic acid is subjected to restriction endonuclease digestion. For example, one or more nucleic acid molecule or fragment thereof of the present invention can be used as a probe in accordance with the above-described polymorphic methods. The polymorphism obtained in this approach can then be cloned to identify the mutation at the coding region which alters the protein's structure or regulatory region of the gene which affects its expression level.

In an aspect of the present invention, one or more of the nucleic molecules of the present invention are used to determine the level (i.e., the concentration of mRNA in a sample, etc.) in a plant (preferably maize or soybean) or pattern (i.e., the kinetics of expression, rate of decomposition, stability profile, etc.) of the expression of a protein encoded in part or whole by one or more of the nucleic acid molecule of the present invention (collectively, the “Expression Response” of a cell or tissue). As used herein, the Expression Response manifested by a cell or tissue is said to be “altered” if it differs from the Expression Response of cells or tissues of plants not exhibiting the phenotype. To determine whether a Expression Response is altered, the Expression Response manifested by the cell or tissue of the plant exhibiting the phenotype is compared with that of a similar cell or tissue sample of a plant not exhibiting the phenotype. As will be appreciated, it is not necessary to re-determine the Expression Response of the cell or tissue sample of plants not exhibiting the phenotype each time such a comparison is made; rather, the Expression Response of a particular plant may be compared with previously obtained values of normal plants. As used herein, the phenotype of the organism is any of one or more characteristics of an organism (e.g. disease resistance, pest tolerance, environmental tolerance such as tolerance to abiotic stress, male sterility, quality improvement or yield etc.). A change in genotype or phenotype may be transient or permanent. Also as used herein, a tissue sample is any sample that comprises more than one cell. In a preferred aspect, a tissue sample comprises cells that share a common characteristic (e.g. derived from root, seed, flower, leaf, stem or pollen etc.).

In one aspect of the present invention, an evaluation can be conducted to determine whether a particular mRNA molecule is present. One or more of the nucleic acid molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention are utilized to detect the presence or quantity of the mRNA species. Such molecules are then incubated with cell or tissue extracts of a plant under conditions sufficient to permit nucleic acid hybridization. The detection of double-stranded probe-mRNA hybrid molecules is indicative of the presence of the mRNA; the amount of such hybrid formed is proportional to the amount of mRNA. Thus, such probes may be used to ascertain the level and extent of the mRNA production in a plant's cells or tissues. Such nucleic acid hybridization may be conducted under quantitative conditions (thereby providing a numerical value of the amount of the mRNA present). Alternatively, the assay may be conducted as a qualitative assay that indicates either that the mRNA is present, or that its level exceeds a user set, predefined value.

A principle of in situ hybridization is that a labeled, single-stranded nucleic acid probe will hybridize to a complementary strand of cellular DNA or RNA and, under the appropriate conditions, these molecules will form a stable hybrid. When nucleic acid hybridization is combined with histological techniques, specific DNA or RNA sequences can be identified within a single cell. An advantage of in situ hybridization over more conventional techniques for the detection of nucleic acids is that it allows an investigator to determine the precise spatial population (Angerer et al., Dev. Biol. 101:477-484 (1984), the entirety of which is herein incorporated by reference; Angerer et al., Dev. Biol. 112:157-166 (1985), the entirety of which is herein incorporated by reference; Dixon et al., EMBO J. 10:1317-1324 (1991), the entirety of which is herein incorporated by reference). In situ hybridization may be used to measure the steady-state level of RNA accumulation. It is a sensitive technique and RNA sequences present in as few as 5-10 copies per cell can be detected (Hardin et al., J. Mol. Biol. 202:417-431 (1989), the entirety of which is herein incorporated by reference). A number of protocols have been devised for in situ hybridization, each with tissue preparation, hybridization and washing conditions (Meyerowitz, Plant Mol. Biol. Rep. 5:242-250 (1987), the entirety of which is herein incorporated by reference; Cox and Goldberg, In: Plant Molecular Biology: A Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford (1988), the entirety of which is herein incorporated by reference; Raikhel et al., In situ RNA hybridization in plant tissues, In: Plant Molecular Biology Manual, vol. B9:1-32, Kluwer Academic Publisher, Dordrecht, Belgium (1989), the entirety of which is herein incorporated by reference).

In situ hybridization also allows for the localization of proteins within a tissue or cell (Wilkinson, In Situ Hybridization, Oxford University Press, Oxford (1992), the entirety of which is herein incorporated by reference; Langdale, In Situ Hybridization In: The Maize Handbook, Freeling and Walbot (eds.), pp 165-179, Springer-Verlag, New York (1994), the entirety of which is herein incorporated by reference). It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the level or pattern of a sucrose pathway protein or mRNA thereof by in situ hybridization.

Fluorescent in situ hybridization allows the localization of a particular DNA sequence along a chromosome which is useful, among other uses, for gene mapping, following chromosomes in hybrid lines or detecting chromosomes with translocations, transversions or deletions. In situ hybridization has been used to identify chromosomes in several plant species (Griffor et al., Plant Mol. Biol. 17:101-109 (1991), the entirety of which is herein incorporated by reference; Gustafson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:1899-1902 (1990), herein incorporated by reference; Mukai and Gill, Genome 34:448-452 (1991), the entirety of which is herein incorporated by reference; Schwarzacher and Heslop-Harrison, Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet. 66:313-316 (1991), the entirety of which is herein incorporated by reference; Parra and Windle, Nature Genetics 5:17-21 (1993), the entirety of which is herein incorporated by reference). It is understood that the nucleic acid molecules of the present invention may be used as probes or markers to localize sequences along a chromosome.

Another method to localize the expression of a molecule is tissue printing. Tissue printing provides a way to screen, at the same time on the same membrane many tissue sections from different plants or different developmental stages. Tissue-printing procedures utilize films designed to immobilize proteins and nucleic acids. In essence, a freshly cut section of a tissue is pressed gently onto nitrocellulose paper, nylon membrane or polyvinylidene difluoride membrane. Such membranes are commercially available (e.g. Millipore, Bedford, Mass. U.S.A.). The contents of the cut cell transfer onto the membrane and the contents and are immobilized to the membrane. The immobilized contents form a latent print that can be visualized with appropriate probes. When a plant tissue print is made on nitrocellulose paper, the cell walls leave a physical print that makes the anatomy visible without further treatment (Varner and Taylor, Plant Physiol. 91:31-33 (1989), the entirety of which is herein incorporated by reference).

Tissue printing on substrate films is described by Daoust, Exp. Cell Res. 12:203-211 (1957), the entirety of which is herein incorporated by reference, who detected amylase, protease, ribonuclease and deoxyribonuclease in animal tissues using starch, gelatin and agar films. These techniques can be applied to plant tissues (Yomo and Taylor, Planta 112:35-43 (1973); the entirety of which is herein incorporated by reference; Harris and Chrispeels, Plant Physiol. 56:292-299 (1975), the entirety of which is herein incorporated by reference). Advances in membrane technology have increased the range of applications of Daoust's tissue-printing techniques allowing (Cassab and Varner, J. Cell. Biol. 105:2581-2588 (1987), the entirety of which is herein incorporated by reference) the histochemical localization of various plant enzymes and deoxyribonuclease on nitrocellulose paper and nylon (Spruce et al., Phytochemistry 26:2901-2903 (1987), the entirety of which is herein incorporated by reference; Barres et al., Neuron 5:527-544 (1990), the entirety of which is herein incorporated by reference; Reid and Pont-Lezica, Tissue Printing: Tools for the Study of Anatomy, Histochemistry and Gene Expression, Academic Press, New York, N.Y. (1992), the entirety of which is herein incorporated by reference; Reid et al., Plant Physiol. 93:160-165 (1990), the entirety of which is herein incorporated by reference; Ye et al., Plant J. 1:175-183 (1991), the entirety of which is herein incorporated by reference).

It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the presence or quantity of a sucrose pathway protein by tissue printing.

Further it is also understood that any of the nucleic acid molecules of the present invention may be used as marker nucleic acids and or probes in connection with methods that require probes or marker nucleic acids. As used herein, a probe is an agent that is utilized to determine an attribute or feature (e.g. presence or absence, location, correlation, etc.) of a molecule, cell, tissue or plant. As used herein, a marker nucleic acid is a nucleic acid molecule that is utilized to determine an attribute or feature (e.g., presence or absence, location, correlation, etc.) or a molecule, cell, tissue or plant.

A microarray-based method for high-throughput monitoring of plant gene expression may be utilized to measure gene-specific hybridization targets. This ‘chip’-based approach involves using microarrays of nucleic acid molecules as gene-specific hybridization targets to quantitatively measure expression of the corresponding plant genes (Schena et al., Science 270:467-470 (1995), the entirety of which is herein incorporated by reference; Shalon, Ph.D. Thesis, Stanford University (1996), the entirety of which is herein incorporated by reference). Every nucleotide in a large sequence can be queried at the same time. Hybridization can be used to efficiently analyze nucleotide sequences.

Several microarray methods have been described. One method compares the sequences to be analyzed by hybridization to a set of oligonucleotides representing all possible subsequences (Bains and Smith, J. Theor. Biol. 135:303-307 (1989), the entirety of which is herein incorporated by reference). A second method hybridizes the sample to an array of oligonucleotide or cDNA molecules. An array consisting of oligonucleotides complementary to subsequences of a target sequence can be used to determine the identity of a target sequence, measure its amount and detect differences between the target and a reference sequence. Nucleic acid molecules microarrays may also be screened with protein molecules or fragments thereof to determine nucleic acid molecules that specifically bind protein molecules or fragments thereof.

The microarray approach may be used with polypeptide targets (U.S. Pat. Nos. 5,445,934; 5,143,854; 5,079,600; 4,923,901, all of which are herein incorporated by reference in their entirety). Essentially, polypeptides are synthesized on a substrate (microarray) and these polypeptides can be screened with either protein molecules or fragments thereof or nucleic acid molecules in order to screen for either protein molecules or fragments thereof or nucleic acid molecules that specifically bind the target polypeptides. (Fodor et al., Science 251:767-773 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules or protein or fragments thereof of the present invention may be utilized in a microarray based method.

In a preferred embodiment of the present invention microarrays may be prepared that comprise nucleic acid molecules where such nucleic acid molecules encode at least one, preferably at least two, more preferably at least three or preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, preferably at least ten, preferably at least eleven, preferably at least twelve, preferably at least thirteen, preferably at least fourteen preferably at least fifteen sucrose pathway enzymes. In a preferred embodiment the nucleic acid molecules are selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or fragment thereof.

Site directed mutagenesis may be utilized to modify nucleic acid sequences, particularly as it is a technique that allows one or more of the amino acids encoded by a nucleic acid molecule to be altered (e.g. a threonine to be replaced by a methionine). Three basic methods for site directed mutagenesis are often employed. These are cassette mutagenesis (Wells et al., Gene 34:315-323 (1985), the entirety of which is herein incorporated by reference), primer extension (Gilliam et al., Gene 12:129-137 (1980), the entirety of which is herein incorporated by reference; Zoller and Smith, Methods Enzymol. 100:468-500 (1983), the entirety of which is herein incorporated by reference; Dalbadie-McFarland et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413 (1982), the entirety of which is herein incorporated by reference) and methods based upon PCR (Scharf et al., Science 233:1076-1078 (1986), the entirety of which is herein incorporated by reference; Higuchi et al, Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is herein incorporated by reference). Site directed mutagenesis approaches are also described in European Patent 0 385 962, the entirety of which is herein incorporated by reference; European Patent 0 359 472, the entirety of which is herein incorporated by reference; and PCT Patent Application WO 93/07278, the entirety of which is herein incorporated by reference.

Site directed mutagenesis strategies have been applied to plants for both in vitro as well as in vivo site directed mutagenesis (Lanz et al., J. Biol. Chem. 266:9971-9976 (1991), the entirety of which is herein incorporated by reference; Kovgan and Zhdanov, Biotekhnologiya 5:148-154; No. 207160n, Chemical Abstracts 110:225 (1989), the entirety of which is herein incorporated by reference; Ge et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:4037-4041 (1989), the entirety of which is herein incorporated by reference; Zhu et al., J. Biol. Chem. 271:18494-18498 (1996), the entirety of which is herein incorporated by reference; Chu et al., Biochemistry 33:6150-6157 (1994), the entirety of which is herein incorporated by reference; Small et al., EMBO J. 11:1291-1296 (1992), the entirety of which is herein incorporated by reference; Cho et al., Mol. Biotechnol. 8:13-16 (1997), the entirety of which is herein incorporated by reference; Kita et al., J. Biol. Chem. 271:26529-26535 (1996), the entirety of which is herein incorporated by reference, Jin et al., Mol. Microbiol. 7:555-562 (1993), the entirety of which is herein incorporated by reference; Hatfield and Vierstra, J. Biol. Chem. 267:14799-14803 (1992), the entirety of which is herein incorporated by reference; Zhao et al., Biochemistry 31:5093-5099 (1992), the entirety of which is herein incorporated by reference).

Any of the nucleic acid molecules of the present invention may either be modified by site directed mutagenesis or used as, for example, nucleic acid molecules that are used to target other nucleic acid molecules for modification. It is understood that mutants with more than one altered nucleotide can be constructed using techniques that practitioners are familiar with such as isolating restriction fragments and ligating such fragments into an expression vector (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989)).

Sequence-specific DNA-binding proteins play a role in the regulation of transcription. The isolation of recombinant cDNAs encoding these proteins facilitates the biochemical analysis of their structural and functional properties. Genes encoding such DNA-binding proteins have been isolated using classical genetics (Vollbrecht et al., Nature 350: 241-243 (1991), the entirety of which is herein incorporated by reference) and molecular biochemical approaches, including the screening of recombinant cDNA libraries with antibodies (Landschulz et al., Genes Dev. 2:786-800 (1988), the entirety of which is herein incorporated by reference) or DNA probes (Bodner et al., Cell 55:505-518 (1988), the entirety of which is herein incorporated by reference). In addition, an in situ screening procedure has been used and has facilitated the isolation of sequence-specific DNA-binding proteins from various plant species (Gilmartin et al., Plant Cell 4:839-849 (1992), the entirety of which is herein incorporated by reference; Schindler et al., EMBO J. 11:1261- 1273 (1992), the entirety of which is herein incorporated by reference). An in situ screening protocol does not require the purification of the protein of interest (Vinson et al., Genes Dev. 2:801-806 (1988), the entirety of which is herein incorporated by reference; Singh et al., Cell 52:415-423 (1988), the entirety of which is herein incorporated by reference).

Two steps may be employed to characterize DNA-protein interactions. The first is to identify promoter fragments that interact with DNA-binding proteins, to titrate binding activity, to determine the specificity of binding and to determine whether a given DNA-binding activity can interact with related DNA sequences (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2^(nd) edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Electrophoretic mobility-shift assay is a widely used assay. The assay provides a rapid and sensitive method for detecting DNA-binding proteins based on the observation that the mobility of a DNA fragment through a nondenaturing, low-ionic strength polyacrylamide gel is retarded upon association with a DNA-binding protein (Fried and Crother, Nucleic Acids Res. 9:6505-6525 (1981), the entirety of which is herein incorporated by reference). When one or more specific binding activities have been identified, the exact sequence of the DNA bound by the protein may be determined. Several procedures for characterizing protein/DNA-binding sites are used, including methylation and ethylation interference assays (Maxam and Gilbert, Methods Enzymol. 65:499-560 (1980), the entirety of which is herein incorporated by reference; Wissman and Hillen, Methods Enzymol. 208:365-379 (1991), the entirety of which is herein incorporated by reference), footprinting techniques employing DNase I (Galas and Schmitz, Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which is herein incorporated by reference), 1,10-phenanthroline-copper ion methods (Sigman et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference) and hydroxyl radicals methods (Dixon et al, Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention may be utilized to identify a protein or fragment thereof that specifically binds to a nucleic acid molecule of the present invention. It is also understood that one or more of the protein molecules or fragments thereof of the present invention may be utilized to identify a nucleic acid molecule that specifically binds to it.

A two-hybrid system is based on the fact that many cellular functions are carried out by proteins, such as transcription factors, that interact (physically) with one another. Two-hybrid systems have been used to probe the function of new proteins (Chien et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9578-9582 (1991) the entirety of which is herein incorporated by reference; Durfee et al., Genes Dev. 7:555-569 (1993) the entirety of which is herein incorporated by reference; Choi et al., Cell 78:499-512 (1994), the entirety of which is herein incorporated by reference; Kranz et al., Genes Dev. 8:313-327 (1994), the entirety of which is herein incorporated by reference).

Interaction mating techniques have facilitated a number of two-hybrid studies of protein-protein interaction. Interaction mating has been used to examine interactions between small sets of tens of proteins (Finley and Brent, Proc. Natl. Acad. Sci. (U.S.A.) 91:12098-12984 (1994), the entirety of which is herein incorporated by reference), larger sets of hundreds of proteins (Bendixen et al., Nucl. Acids Res. 22:1778-1779 (1994), the entirety of which is herein incorporated by reference) and to comprehensively map proteins encoded by a small genome (Bartel et al., Nature Genetics 12:72-77 (1996), the entirety of which is herein incorporated by reference). This technique utilizes proteins fused to the DNA-binding domain and proteins fused to the activation domain. They are expressed in two different haploid yeast strains of opposite mating type and the strains are mated to determine if the two proteins interact. Mating occurs when haploid yeast strains come into contact and result in the fusion of the two haploids into a diploid yeast strain. An interaction can be determined by the activation of a two-hybrid reporter gene in the diploid strain. An advantage of this technique is that it reduces the number of yeast transformations needed to test individual interactions. It is understood that the protein-protein interactions of protein or fragments thereof of the present invention may be investigated using the two-hybrid system and that any of the nucleic acid molecules of the present invention that encode such proteins or fragments thereof may be used to transform yeast in the two-hybrid system.

(a) Plant Constructs and Plant Transformants

One or more of the nucleic acid molecules of the present invention may be used in plant transformation or transfection. Exogenous genetic material may be transferred into a plant cell and the plant cell regenerated into a whole, fertile or sterile plant. Exogenous genetic material is any genetic material, whether naturally occurring or otherwise, from any source that is capable of being inserted into any organism. Such genetic material may be transferred into either monocotyledons and dicotyledons including, but not limited to maize (pp 63-69), soybean (pp 50-60), Arabidopsis (p 45), phaseolus (pp 47-49), peanut (pp 49-50), alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79), oat (pp 80-81), sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p85), fescue (p 85), perennial ryegrass (p 86), sugarcane (p87), cranberry (p101), papaya (pp 101-102), banana (p 103), banana (p 103), muskmelon (p 104), apple (p 104), cucumber (p 105), dendrobium (p 109), gladiolus (p 110), chrysanthemum (p 110), liliacea (p 111), cotton (pp113-114), eucalyptus (p 115), sunflower (p 118), canola (p 118), turfgrass (p121), sugarbeet (p 122), coffee (p 122) and dioscorea (p 122), (Christou, In: Particle Bombardment for Genetic Engineering of Plants, Biotechnology Intelligence Unit, Academic Press, San Diego, Calif. (1996), the entirety of which is herein incorporated by reference).

Transfer of a nucleic acid that encodes for a protein can result in overexpression of that protein in a transformed cell or transgenic plant. One or more of the proteins or fragments thereof encoded by nucleic acid molecules of the present invention may be overexpressed in a transformed cell or transformed plant. Particularly, any of the sucrose pathway proteins or fragments thereof may be overexpressed in a transformed cell or transgenic plant. Such overexpression may be the result of transient or stable transfer of the exogenous genetic material.

Exogenous genetic material may be transferred into a plant cell and the plant cell by the use of a DNA vector or construct designed for such a purpose. Design of such a vector is generally within the skill of the art (See, Plant Molecular Biology: A Laboratory Manual, Clark (ed.), Springier, N. Y. (1997), the entirety of which is herein incorporated by reference).

A construct or vector may include a plant promoter to express the protein or protein fragment of choice. A number of promoters which are active in plant cells have been described in the literature. These include the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987), the entirety of which is herein incorporated by reference), the octopine synthase (OCS) promoter (which are carried on tumor-inducing plasmids of Agrobacterium tumefaciens), the caulimovirus promoters such as the cauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), the entirety of which is herein incorporated by reference) and the CAMV 35S promoter (Odell et al., Nature 313:810-812 (1985), the entirety of which is herein incorporated by reference), the figwort mosaic virus 35S-promoter, the light-inducible promoter from the small subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:4144-4148 (1990), the entirety of which is herein incorporated by reference), the R gene complex promoter (Chandler et al., The Plant Cell 1:1175-1183 (1989), the entirety of which is herein incorporated by reference) and the chlorophyll a/b binding protein gene promoter, etc. These promoters have been used to create DNA constructs which have been expressed in plants; see, e.g., PCT publication WO 84/02913, herein incorporated by reference in its entirety.

Promoters which are known or are found to cause transcription of DNA in plant cells can be used in the present invention. Such promoters may be obtained from a variety of sources such as plants and plant viruses. It is preferred that the particular promoter selected should be capable of causing sufficient expression to result in the production of an effective amount of the sucrose pathway protein to cause the desired phenotype. In addition to promoters that are known to cause transcription of DNA in plant cells, other promoters may be identified for use in the current invention by screening a plant cDNA library for genes which are selectively or preferably expressed in the target tissues or cells.

For the purpose of expression in source tissues of the plant, such as the leaf, seed, root or stem, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. For this purpose, one may choose from a number of promoters for genes with tissue- or cell-specific or -enhanced expression. Examples of such promoters reported in the literature include the chloroplast glutamine synthetase GS2 promoter from pea (Edwards et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:3459-3463 (1990), herein incorporated by reference in its entirety), the chloroplast fructose-1,6-biphosphatase (FBPase) promoter from wheat (Lloyd et al., Mol. Gen. Genet. 225:209-216 (1991), herein incorporated by reference in its entirety), the nuclear photosynthetic ST-LS1 promoter from potato (Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated by reference in its entirety), the serine/threonine kinase (PAL) promoter and the glucoamylase (CHS) promoter from Arabidopsis thaliana. Also reported to be active in photosynthetically active tissues are the ribulose-1,5-bisphosphate carboxylase (RbcS) promoter from eastern larch (Larix laricina), the promoter for the cab gene, cab6, from pine (Yamamoto et al., Plant Cell Physiol. 35:773-778 (1994), herein incorporated by reference in its entirety), the promoter for the Cab-1 gene from wheat (Fejes et al., Plant Mol. Biol. 15:921-932 (1990), herein incorporated by reference in its entirety), the promoter for the CAB-1 gene from spinach (Lubberstedt et al., Plant Physiol. 104:997-1006 (1994), herein incorporated by reference in its entirety), the promoter for the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981 (1992), the entirety of which is herein incorporated by reference), the pyruvate, orthophosphate dikinase (PPDK) promoter from maize (Matsuoka et al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590 (1993), herein incorporated by reference in its entirety), the promoter for the tobacco Lhcb1*2 gene (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997), herein incorporated by reference in its entirety), the Arabidopsis thaliana SUC2 sucrose-H+ symporter promoter (Truernit et al., Planta. 196:564-570 (1995), herein incorporated by reference in its entirety) and the promoter for the thylakoid membrane proteins from spinach (psaD, psaF, psaE, PC, FNR, atpC, atpD, cab, rbcS). Other promoters for the chlorophyll a/b-binding proteins may also be utilized in the present invention, such as the promoters for LhcB gene and PsbP gene from white mustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229 (1995), the entirety of which is herein incorporated by reference).

For the purpose of expression in sink tissues of the plant, such as the tuber of the potato plant, the fruit of tomato, or the seed of maize, wheat, rice and barley, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. A number of promoters for genes with tuber-specific or -enhanced expression are known, including the class I patatin promoter (Bevan et al., EMBO J. 8:1899-1906 (1986); Jefferson et al., Plant Mol. Biol. 14:995-1006 (1990), both of which are herein incorporated by reference in its entirety), the promoter for the potato tuber ADPGPP genes, both the large and small subunits, the sucrose synthase promoter (Salanoubat and Belliard, Gene. 60:47-56 (1987), Salanoubat and Belliard, Gene. 84:181-185 (1989), both of which are incorporated by reference in their entirety), the promoter for the major tuber proteins including the 22 kd protein complexes and proteinase inhibitors (Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated by reference in its entirety), the promoter for the granule bound starch synthase gene (GBSS) (Visser et al., Plant Mol. Biol. 17:691-699 (1991), herein incorporated by reference in its entirety) and other class I and II patatins promoters (Koster-Topfer et al., Mol Gen Genet. 219:390-396 (1989); Mignery et al., Gene. 62:27-44 (1988), both of which are herein incorporated by reference in their entirety).

Other promoters can also be used to express a sucrose pathway protein or fragment thereof in specific tissues, such as seeds or fruits. The promoter for β-conglycinin (Chen et al., Dev. Genet. 10: 112-122 (1989), herein incorporated by reference in its entirety) or other seed-specific promoters such as the napin and phaseolin promoters, can be used. The zeins are a group of storage proteins found in maize endosperm. Genomic clones for zein genes have been isolated (Pedersen et al., Cell 29:1015-1026 (1982), herein incorporated by reference in its entirety) and the promoters from these clones, including the 15 kD, 16 kD, 19 kD, 22 kD, 27 kD and genes, could also be used. Other promoters known to function, for example, in maize include the promoters for the following genes: waxy, Brittle, Shrunken 2, Branching enzymes I and II, starch synthases, debranching enzymes, oleosins, glutelins and sucrose synthases. A particularly preferred promoter for maize endosperm expression is the promoter for the glutelin gene from rice, more particularly the Osgt-1 promoter (Zheng et al., Mol. Cell Biol. 13:5829-5842 (1993), herein incorporated by reference in its entirety). Examples of promoters suitable for expression in wheat include those promoters for the ADPglucose pyrosynthase (ADPGPP) subunits, the granule bound and other starch synthase, the branching and debranching enzymes, the embryogenesis-abundant proteins, the gliadins and the glutenins. Examples of such promoters in rice include those promoters for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases and the glutelins. A particularly preferred promoter is the promoter for rice glutelin, Osgt-1. Examples of such promoters for barley include those for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases, the hordeins, the embryo globulins and the aleurone specific proteins.

Root specific promoters may also be used. An example of such a promoter is the promoter for the acid chitinase gene (Samac et al., Plant Mol. Biol. 25:587-596 (1994), the entirety of which is herein incorporated by reference). Expression in root tissue could also be accomplished by utilizing the root specific subdomains of the CaMV35S promoter that have been identified (Lam et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:7890-7894 (1989), herein incorporated by reference in its entirety). Other root cell specific promoters include those reported by Conkling et al. (Conkling et al., Plant Physiol. 93:1203-1211 (1990), the entirety of which is herein incorporated by reference).

Additional promoters that may be utilized are described, for example, in U.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147; 5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441; 5,633,435; and 4,633,436, all of which are herein incorporated in their entirety. In addition, a tissue specific enhancer may be used (Fromm et al., The Plant Cell 1:977-984 (1989), the entirety of which is herein incorporated by reference).

Constructs or vectors may also include with the coding region of interest a nucleic acid sequence that acts, in whole or in part, to terminate transcription of that region. For example, such sequences have been isolated including the Tr7 3′ sequence and the NOS 3′ sequence (Ingelbrecht et al., The Plant Cell 1:671-680 (1989), the entirety of which is herein incorporated by reference; Bevan et al., Nucleic Acids Res. 11:369-385 (1983), the entirety of which is herein incorporated by reference), or the like.

A vector or construct may also include regulatory elements. Examples of such include the Adh intron 1 (Callis et al., Genes and Develop. 1:1183-1200 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase intron (Vasil et al., Plant Physiol. 91:1575-1579 (1989), the entirety of which is herein incorporated by reference) and the TMV omega element (Gallie et al., The Plant Cell 1:301-311 (1989), the entirety of which is herein incorporated by reference). These and other regulatory elements may be included when appropriate.

A vector or construct may also include a selectable marker. Selectable markers may also be used to select for plants or plant cells that contain the exogenous genetic material. Examples of such include, but are not limited to, a neo gene (Potrykus et al., Mol. Gen. Genet. 199:183-188 (1985), the entirety of which is herein incorporated by reference) which codes for kanamycin resistance and can be selected for using kanamycin, G418, etc.; a bar gene which codes for bialaphos resistance; a mutant EPSP synthase gene (Hinchee et al., Bio/Technology 6:915-922 (1988), the entirety of which is herein incorporated by reference) which encodes glyphosate resistance; a nitrilase gene which confers resistance to bromoxynil (Stalker et al., J. Biol. Chem. 263:6310-6314 (1988), the entirety of which is herein incorporated by reference); a mutant acetolactate synthase gene (ALS) which confers imidazolinone or sulphonylurea resistance (European Patent Application 154,204 (Sep. 11, 1985), the entirety of which is herein incorporated by reference); and a methotrexate resistant DHFR gene (Thillet et al., J. Biol. Chem. 263:12500-12508 (1988), the entirety of which is herein incorporated by reference).

A vector or construct may also include a transit peptide. Incorporation of a suitable chloroplast transit peptide may also be employed (European Patent Application Publication Number 0218571, the entirety of which is herein incorporated by reference). Translational enhancers may also be incorporated as part of the vector DNA. DNA constructs could contain one or more 5′ non-translated leader sequences which may serve to enhance expression of the gene products from the resulting mRNA transcripts. Such sequences may be derived from the promoter selected to express the gene or can be specifically modified to increase translation of the mRNA. Such regions may also be obtained from viral RNAs, from suitable eukaryotic genes, or from a synthetic gene sequence. For a review of optimizing expression of transgenes, see Koziel et al., Plant Mol. Biol. 32:393-405 (1996), the entirety of which is herein incorporated by reference.

A vector or construct may also include a screenable marker. Screenable markers may be used to monitor expression. Exemplary screenable markers include a β-glucuronidase or uidA gene (GUS) which encodes an enzyme for which various chromogenic substrates are known (Jefferson, Plant Mol. Biol, Rep. 5:387-405 (1987), the entirety of which is herein incorporated by reference; Jefferson et al., EMBO J. 6:3901-3907 (1987), the entirety of which is herein incorporated by reference); an R-locus gene, which encodes a product that regulates the production of anthocyanin pigments (red color) in plant tissues (Dellaporta et al., Stadler Symposium 11:263-282 (1988), the entirety of which is herein incorporated by reference); a β-lactamase gene (Sutcliffe et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:3737-3741 (1978), the entirety of which is herein incorporated by reference), a gene which encodes an enzyme for which various chromogenic substrates are known (e.g., PADAC, a chromogenic cephalosporin); a luciferase gene (Ow et al., Science 234:856-859 (1986), the entirety of which is herein incorporated by reference); a xylE gene (Zukowsky et al., Proc. Natl. Acad. Sci. (U.S.A.) 80:1101-1105 (1983), the entirety of which is herein incorporated by reference) which encodes a catechol diozygenase that can convert chromogenic catechols; an α-amylase gene (Ikatu et al., Bio/Technol. 8:241-242 (1990), the entirety of which is herein incorporated by reference); a tyrosinase gene (Katz et al., J. Gen. Microbiol. 129:2703-2714 (1983), the entirety of which is herein incorporated by reference) which encodes an enzyme capable of oxidizing tyrosine to DOPA and dopaquinone which in turn condenses to melanin; an α-galactosidase, which will turn a chromogenic α-galactose substrate.

Included within the terms “selectable or screenable marker genes” are also genes which encode a secretable marker whose secretion can be detected as a means of identifying or selecting for transformed cells. Examples include markers which encode a secretable antigen that can be identified by antibody interaction, or even secretable enzymes which can be detected catalytically. Secretable proteins fall into a number of classes, including small, diffusible proteins which are detectable, (e.g., by ELISA), small active enzymes which are detectable in extracellular solution (e.g. α-amylase, β-lactamase, phosphinothricin transferase), or proteins which are inserted or trapped in the cell wall (such as proteins which include a leader sequence such as that found in the expression unit of extension or tobacco PR-S). Other possible selectable and/or screenable marker genes will be apparent to those of skill in the art.

There are many methods for introducing transforming nucleic acid molecules into plant cells. Suitable methods are believed to include virtually any method by which nucleic acid molecules may be introduced into a cell, such as by Agrobacterium infection or direct delivery of nucleic acid molecules such as, for example, by PEG-mediated transformation, by electroporation or by acceleration of DNA coated particles, etc (Potrykus, Ann. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225 (1991), the entirety of which is herein incorporated by reference; Vasil, Plant Mol. Biol. 25:925-937 (1994), the entirety of which is herein incorporated by reference). For example, electroporation has been used to transform maize protoplasts (Fromm et al., Nature 312:791-793 (1986), the entirety of which is herein incorporated by reference).

Other vector systems suitable for introducing transforming DNA into a host plant cell include but are not limited to binary artificial chromosome (BIBAC) vectors (Hamilton et al., Gene 200:107-116 (1997), the entirety of which is herein incorporated by reference); and transfection with RNA viral vectors (Della-Cioppa et al., Ann. N.Y. Acad. Sci. (1996), 792 (Engineering Plants for Commercial Products and Applications), 57-61, the entirety of which is herein incorporated by reference). Additional vector systems also include plant selectable YAC vectors such as those described in Mullen et al., Molecular Breeding 4:449-457 (1988), the entireity of which is herein incorporated by reference).

Technology for introduction of DNA into cells is well known to those of skill in the art. Four general methods for delivering a gene into cells have been described: (1) chemical methods (Graham and van der Eb, Virology 54:536-539 (1973), the entirety of which is herein incorporated by reference); (2) physical methods such as microinjection (Capecchi, Cell 22:479-488 (1980), the entirety of which is herein incorporated by reference), electroporation (Wong and Neumann, Biochem. Biophys. Res. Commun. 107:584-587 (1982); Fromm et al., Proc. Natl. Acad. Sci. (U.S.A.) 82:5824-5828 (1985); U.S. Pat. No. 5,384,253, all of which are herein incorporated in their entirety); and the gene gun (Johnston and Tang, Methods Cell Biol. 43:353-365 (1994), the entirety of which is herein incorporated by reference); (3) viral vectors (Clapp, Clin. Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096 (1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all of which are herein incorporated in their entirety); and (4) receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther. 3:147-154 (1992), Wagner et al., Proc. Natl. Acad. Sci. (USA) 89:6099-6103 (1992), both of which are incorporated by reference in their entirety).

Acceleration methods that may be used include, for example, microprojectile bombardment and the like. One example of a method for delivering transforming nucleic acid molecules to plant cells is microprojectile bombardment. This method has been reviewed by Yang and Christou (eds.), Particle Bombardment Technology for Gene Transfer, Oxford Press, Oxford, England (1994), the entirety of which is herein incorporated by reference). Non-biological particles (microprojectiles) that may be coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, gold, platinum and the like.

A particular advantage of microprojectile bombardment, in addition to it being an effective means of reproducibly transforming monocots, is that neither the isolation of protoplasts (Cristou et al., Plant Physiol. 87:671-674 (1988), the entirety of which is herein incorporated by reference) nor the susceptibility of Agrobacterium infection are required. An illustrative embodiment of a method for delivering DNA into maize cells by acceleration is a biolistics α-particle delivery system, which can be used to propel particles coated with DNA through a screen, such as a stainless steel or Nytex screen, onto a filter surface covered with corn cells cultured in suspension. Gordon-Kamm et al., describes the basic procedure for coating tungsten particles with DNA (Gordon-Kamm et al., Plant Cell 2:603-618 (1990), the entirety of which is herein incorporated by reference). The screen disperses the tungsten nucleic acid particles so that they are not delivered to the recipient cells in large aggregates. A particle delivery system suitable for use with the present invention is the helium acceleration PDS-1000/He gun is available from Bio-Rad Laboratories (Bio-Rad, Hercules, Calif.)(Sanford et al., Technique 3:3-16 (1991), the entirety of which is herein incorporated by reference).

For the bombardment, cells in suspension may be concentrated on filters. Filters containing the cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the gun and the cells to be bombarded.

Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the acceleration device and the cells to be bombarded. Through the use of techniques set forth herein one may obtain up to 1000 or more foci of cells transiently expressing a marker gene. The number of cells in a focus which express the exogenous gene product 48 hours post-bombardment often range from one to ten and average one to three.

In bombardment transformation, one may optimize the pre-bombardment culturing conditions and the bombardment parameters to yield the maximum numbers of stable transformants. Both the physical and biological parameters for bombardment are important in this technology. Physical factors are those that involve manipulating the DNA/microprojectile precipitate or those that affect the flight and velocity of either the macro- or microprojectiles. Biological factors include all steps involved in manipulation of cells before and immediately after bombardment, the osmotic adjustment of target cells to help alleviate the trauma associated with bombardment and also the nature of the transforming DNA, such as linearized DNA or intact supercoiled plasmids. It is believed that pre-bombardment manipulations are especially important for successful transformation of immature embryos.

In another alternative embodiment, plastids can be stably transformed. Methods disclosed for plastid transformation in higher plants include the particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination (Svab et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8526-8530 (1990); Svab and Maliga, Proc. Natl. Acad. Sci. (U.S.A.) 90:913-917 (1993); Staub and Maliga, EMBO J. 12:601-606 (1993); U.S. Pat. Nos. 5,451,513 and 5,545,818, all of which are herein incorporated by reference in their entirety).

Accordingly, it is contemplated that one may wish to adjust various aspects of the bombardment parameters in small scale studies to fully optimize the conditions. One may particularly wish to adjust physical parameters such as gap distance, flight distance, tissue distance and helium pressure. One may also minimize the trauma reduction factors by modifying conditions which influence the physiological state of the recipient cells and which may therefore influence transformation and integration efficiencies. For example, the osmotic state, tissue hydration and the subculture stage or cell cycle of the recipient cells may be adjusted for optimum transformation. The execution of other routine adjustments will be known to those of skill in the art in light of the present disclosure.

Agrobacterium-mediated transfer is a widely applicable system for introducing genes into plant cells because the DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art. See, for example the methods described by Fraley et al., Bio/Technology 3:629-635 (1985) and Rogers et al., Methods Enzymol. 153:253-277 (1987), both of which are herein incorporated by reference in their entirety. Further, the integration of the Ti-DNA is a relatively precise process resulting in few rearrangements. The region of DNA to be transferred is defined by the border sequences and intervening DNA is usually inserted into the plant genome as described (Spielmann et al., Mol. Gen. Genet. 205:34 (1986), the entirety of which is herein incorporated by reference).

Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations as described (Klee et al., In: Plant DNA Infectious Agents, Hohn and Schell (eds.), Springer-Verlag, New York, pp. 179-203 (1985), the entirety of which is herein incorporated by reference. Moreover, technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes and are suitable for present purposes (Rogers et al., Methods Enzymol. 153:253-277 (1987)). In addition, Agrobacterium containing both armed and disarmed Ti genes can be used for the transformations. In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene transfer.

A transgenic plant formed using Agrobacterium transformation methods typically contains a single gene on one chromosome. Such transgenic plants can be referred to as being heterozygous for the added gene. More preferred is a transgenic plant that is homozygous for the added structural gene; i.e., a transgenic plant that contains two added genes, one gene at the same locus on each chromosome of a chromosome pair. A homozygous transgenic plant can be obtained by sexually mating (selfing) an independent segregant transgenic plant that contains a single added gene, germinating some of the seed produced and analyzing the resulting plants produced for the gene of interest.

It is also to be understood that two different transgenic plants can also be mated to produce offspring that contain two independently segregating added, exogenous genes. Selfing of appropriate progeny can produce plants that are homozygous for both added, exogenous genes that encode a polypeptide of interest. Back-crossing to a parental plant and out-crossing with a non-transgenic plant are also contemplated, as is vegetative propagation.

Transformation of plant protoplasts can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation and combinations of these treatments (See, for example, Potrykus et al., Mol. Gen. Genet. 205:193-200 (1986); Lorz et al., Mol. Gen. Genet. 199:178 (1985); Fromm et al., Nature 319:791 (1986); Uchimiya et al., Mol. Gen. Genet. 204:204 (1986); Marcotte et al., Nature 335:454-457 (1988), all of which are herein incorporated by reference in their entirety).

Application of these systems to different plant strains depends upon the ability to regenerate that particular plant strain from protoplasts. Illustrative methods for the regeneration of cereals from protoplasts are described (Fujimura et al., Plant Tissue Culture Letters 2:74 (1985); Toriyama et al., Theor Appl. Genet. 205:34 (1986); Yamada et al., Plant Cell Rep. 4:85 (1986); Abdullah et al., Biotechnolog 4:1087 (1986), all of which are herein incorporated by reference in their entirety).

To transform plant strains that cannot be successfully regenerated from protoplasts, other ways to introduce DNA into intact cells or tissues can be utilized. For example, regeneration of cereals from immature embryos or explants can be effected as described (Vasil, Biotechnology 6:397 (1988), the entirety of which is herein incorporated by reference). In addition, “particle gun” or high-velocity microprojectile technology can be utilized (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference).

Using the latter technology, DNA is carried through the cell wall and into the cytoplasm on the surface of small metal particles as described (Klein et al., Nature 328:70 (1987); Klein et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al., Bio/Technology 6:923 (1988), all of which are herein incorporated by reference in their entirety). The metal particles penetrate through several layers of cells and thus allow the transformation of cells within tissue explants.

Other methods of cell transformation can also be used and include but are not limited to introduction of DNA into plants by direct DNA transfer into pollen (Zhou et al., Methods Enzymol. 101:433 (1983); Hess et al, Intern Rev. Cytol. 107:367 (1987); Luo et al., Plant Mol Biol. Reporter 6:165 (1988), all of which are herein incorporated by reference in their entirety), by direct injection of DNA into reproductive organs of a plant (Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference), or by direct injection of DNA into the cells of immature embryos followed by the rehydration of desiccated embryos (Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entirety of which is herein incorporated by reference).

The regeneration, development and cultivation of plants from single plant protoplast transformants or from various transformed explants is well known in the art (Weissbach and Weissbach, In: Methods for Plant Molecular Biology, Academic Press, San Diego, Calif., (1988), the entirety of which is herein incorporated by reference). This regeneration and growth process typically includes the steps of selection of transformed cells, culturing those individualized cells through the usual stages of embryonic development through the rooted plantlet stage. Transgenic embryos and seeds are similarly regenerated. The resulting transgenic rooted shoots are thereafter planted in an appropriate plant growth medium such as soil.

The development or regeneration of plants containing the foreign, exogenous gene that encodes a protein of interest is well known in the art. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. Otherwise, pollen obtained from the regenerated plants is crossed to seed-grown plants of agronomically important lines. Conversely, pollen from plants of these important lines is used to pollinate regenerated plants. A transgenic plant of the present invention containing a desired polypeptide is cultivated using methods well known to one skilled in the art.

There are a variety of methods for the regeneration of plants from plant tissue. The particular method of regeneration will depend on the starting plant tissue and the particular plant species to be regenerated.

Methods for transforming dicots, primarily by use of Agrobacterium tumefaciens and obtaining transgenic plants have been published for cotton (U.S. Pat. Nos. 5,004,863; 5,159,135; 5,518,908, all of which are herein incorporated by reference in their entirety); soybean (U.S. Pat. Nos. 5,569,834; 5,416,011; McCabe et. al., Biotechnology 6:923 (1988); Christou et al., Plant Physiol. 87:671-674 (1988); all of which are herein incorporated by reference in their entirety); Brassica (U.S. Pat. No. 5,463,174, the entirety of which is herein incorporated by reference); peanut (Cheng et al., Plant Cell Rep. 15:653-657 (1996), McKently et al., Plant Cell Rep. 14:699-703 (1995), all of which are herein incorporated by reference in their entirety); papaya; and pea (Grant et al., Plant Cell Rep. 15:254-258 (1995), the entirety of which is herein incorporated by reference).

Transformation of monocotyledons using electroporation, particle bombardment and Agrobacterium have also been reported. Transformation and plant regeneration have been achieved in asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (USA) 84:5354 (1987), the entirety of which is herein incorporated by reference); barley (Wan and Lemaux, Plant Physiol 104:37 (1994), the entirety of which is herein incorporated by reference); maize (Rhodes et al., Science 240:204 (1988); Gordon-Kamm et al., Plant Cell 2:603-618 (1990); Fromm et al., Bio/Technology 8:833 (1990); Koziel et al., Bio/Technology 11:194 (1993); Armstrong et al., Crop Science 35:550-557 (1995); all of which are herein incorporated by reference in their entirety); oat (Somers et al., Bio/Technology 10:1589 (1992), the entirety of which is herein incorporated by reference); orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988), the entirety of which is herein incorporated by reference); rice (Toriyama et al., Theor Appl. Genet. 205:34 (1986); Part et al., Plant Mol. Biol. 32:1135-1148 (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133-141 (1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al., Plant Cell Rep. 7:379 (1988); Battraw and Hall, Plant Sci. 86:191-202 (1992); Christou et al., Bio/Technology 9:957 (1991), all of which are herein incorporated by reference in their entirety); rye (De la Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference); sugarcane (Bower and Birch, Plant J. 2:409 (1992), the entirety of which is herein incorporated by reference); tall fescue (Wang et al., Bio/Technology 10:691 (1992), the entirety of which is herein incorporated by reference) and wheat (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,631,152, the entirety of which is herein incorporated by reference.)

Assays for gene expression based on the transient expression of cloned nucleic acid constructs have been developed by introducing the nucleic acid molecules into plant cells by polyethylene glycol treatment, electroporation, or particle bombardment (Marcotte et al., Nature 335:454-457 (1988), the entirety of which is herein incorporated by reference; Marcotte et al., Plant Cell 1:523-532 (1989), the entirety of which is herein incorporated by reference; McCarty et al., Cell 66:895-905 (1991), the entirety of which is herein incorporated by reference; Hattori et al., Genes Dev. 6:609-618 (1992), the entirety of which is herein incorporated by reference; Goff et al., EMBO J. 9:2517-2522 (1990), the entirety of which is herein incorporated by reference). Transient expression systems may be used to functionally dissect gene constructs (see generally, Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995)).

Any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a permanent or transient manner in combination with other genetic elements such as vectors, promoters, enhancers etc. Further, any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a manner that allows for overexpression of the protein or fragment thereof encoded by the nucleic acid molecule.

Cosuppression is the reduction in expression levels, usually at the level of RNA, of a particular endogenous gene or gene family by the expression of a homologous sense construct that is capable of transcribing mRNA of the same strandedness as the transcript of the endogenous gene (Napoli et al., Plant Cell 2:279-289 (1990), the entirety of which is herein incorporated by reference; van der Krol et al., Plant Cell 2:291-299 (1990), the entirety of which is herein incorporated by reference). Cosuppression may result from stable transformation with a single copy nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Prolls and Meyer, Plant J. 2:465-475 (1992), the entirety of which is herein incorporated by reference) or with multiple copies of a nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Mittlesten et al., Mol. Gen. Genet. 244:325-330 (1994), the entirety of which is herein incorporated by reference). Genes, even though different, linked to homologous promoters may result in the cosuppression of the linked genes (Vaucheret, C. R. Acad. Sci. III 316:1471-1483 (1993), the entirety of which is herein incorporated by reference).

This technique has, for example, been applied to generate white flowers from red petunia and tomatoes that do not ripen on the vine. Up to 50% of petunia transformants that contained a sense copy of the glucoamylase (CHS) gene produced white flowers or floral sectors; this was as a result of the post-transcriptional loss of mRNA encoding CHS (Flavell, Proc. Natl. Acad. Sci. (U.S.A.) 91:3490-3496 (1994), the entirety of which is herein incorporated by reference); van Blokland et al., Plant J. 6:861-877 (1994), the entirety of which is herein incorporated by reference). Cosuppression may require the coordinate transcription of the transgene and the endogenous gene and can be reset by a developmental control mechanism (Jorgensen, Trends Biotechnol. 8:340-344 (1990), the entirety of which is herein incorporated by reference; Meins and Kunz, In: Gene Inactivation and Homologous Recombination in Plants, Paszkowski (ed.), pp. 335-348, Kluwer Academic, Netherlands (1994), the entirety of which is herein incorporated by reference).

It is understood that one or more of the nucleic acids of the present invention may be introduced into a plant cell and transcribed using an appropriate promoter with such transcription resulting in the cosuppression of an endogenous sucrose pathway protein.

Antisense approaches are a way of preventing or reducing gene function by targeting the genetic material (Mol et al., FEBS Lett. 268:427-430 (1990), the entirety of which is herein incorporated by reference). The objective of the antisense approach is to use a sequence complementary to the target gene to block its expression and create a mutant cell line or organism in which the level of a single chosen protein is selectively reduced or abolished. Antisense techniques have several advantages over other ‘reverse genetic’ approaches. The site of inactivation and its developmental effect can be manipulated by the choice of promoter for antisense genes or by the timing of external application or microinjection. Antisense can manipulate its specificity by selecting either unique regions of the target gene or regions where it shares homology to other related genes (Hiatt et al., In: Genetic Engineering, Setlow (ed.), Vol. 11, New York: Plenum 49-63 (1989), the entirety of which is herein incorporated by reference).

The principle of regulation by antisense RNA is that RNA that is complementary to the target mRNA is introduced into cells, resulting in specific RNA:RNA duplexes being formed by base pairing between the antisense substrate and the target mRNA (Green et al., Annu. Rev. Biochem. 55:569-597 (1986), the entirety of which is herein incorporated by reference). Under one embodiment, the process involves the introduction and expression of an antisense gene sequence. Such a sequence is one in which part or all of the normal gene sequences are placed under a promoter in inverted orientation so that the ‘wrong’ or complementary strand is transcribed into a noncoding antisense RNA that hybridizes with the target mRNA and interferes with its expression (Takayama and Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990), the entirety of which is herein incorporated by reference). An antisense vector is constructed by standard procedures and introduced into cells by transformation, transfection, electroporation, microinjection, infection, etc. The type of transformation and choice of vector will determine whether expression is transient or stable. The promoter used for the antisense gene may influence the level, timing, tissue, specificity, or inducibility of the antisense inhibition.

It is understood that the activity of a sucrose pathway protein in a plant cell may be reduced or depressed by growing a transformed plant cell containing a nucleic acid molecule whose non-transcribed strand encodes a sucrose pathway protein or fragment thereof.

Antibodies have been expressed in plants (Hiatt et al., Nature 342:76-78 (1989), the entirety of which is herein incorporated by reference; Conrad and Fielder, Plant Mol. Biol. 26:1023-1030 (1994), the entirety of which is herein incorporated by reference). Cytoplamsic expression of a scFv (single-chain Fv antibodies) has been reported to delay infection by artichoke mottled crinkle virus. Transgenic plants that express antibodies directed against endogenous proteins may exhibit a physiological effect (Philips et al., EMBO J. 16:4489-4496 (1997), the entirety of which is herein incorporated by reference; Marion-Poll, Trends in Plant Science 2:447-448 (1997), the entirety of which is herein incorporated by reference). For example, expressed anti-abscisic antibodies have been reported to result in a general perturbation of seed development (Philips et al., EMBO J. 16: 4489-4496 (1997)).

Antibodies that are catalytic may also be expressed in plants (abzymes). The principle behind abzymes is that since antibodies may be raised against many molecules, this recognition ability can be directed toward generating antibodies that bind transition states to force a chemical reaction forward (Persidas, Nature Biotechnology 15:1313-1315 (1997), the entirety of which is herein incorporated by reference; Baca et al., Ann. Rev. Biophys. Biomol. Struct. 26:461-493 (1997), the entirety of which is herein incorporated by reference). The catalytic abilities of abzymes may be enhanced by site directed mutagenesis. Examples of abzymes are, for example, set forth in U.S. Pat. Nos: 5,658,753; 5,632,990; 5,631,137; 5,602,015; 5,559,538; 5,576,174; 5,500,358; 5,318,897; 5,298,409; 5,258,289 and 5,194,585, all of which are herein incorporated in their entirety.

It is understood that any of the antibodies of the present invention may be expressed in plants and that such expression can result in a physiological effect. It is also understood that any of the expressed antibodies may be catalytic.

(b) Fungal Constructs and Fungal Transformants

The present invention also relates to a fungal recombinant vector comprising exogenous genetic material. The present invention also relates to a fungal cell comprising a fungal recombinant vector. The present invention also relates to methods for obtaining a recombinant fungal host cell comprising introducing into a fungal host cell exogenous genetic material.

Exogenous genetic material may be transferred into a fungal cell. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention. The fungal recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the fungal host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the fungal host.

The fungal vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the fungal cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the fungal host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the fungal host cell and, furthermore, may be non-encoding or encoding sequences.

For autonomous replication, the vector may further comprise an origin of replication enabling the vector to replicate autonomously in the host cell in question. Examples of origin of replications for use in a yeast host cell are the 2 micron origin of replication and the combination of CEN3 and ARS 1. Any origin of replication may be used which is compatible with the fungal host cell of choice.

The fungal vectors of the present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. The selectable marker may be selected from the group including, but not limited to, amdS (acetamidase), argB (ornithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hygB (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5′-phosphate decarboxylase) and sC (sulfate adenyltransferase) and trpC (anthranilate synthase). Preferred for use in an Aspergillus cell are the amdS and pyrG markers of Aspergillus nidulans or Aspergillus oryzae and the bar marker of Streptomyces hygroscopicus. Furthermore, selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, the entirety of which is herein incorporated by reference. A nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the fungal host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof.

A promoter may be any nucleic acid sequence which shows transcriptional activity in the fungal host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of a nucleic acid construct of the invention in a filamentous fungal host are promoters obtained from the genes encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stable alpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase (glaA), Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulans acetamidase and hybrids thereof. In a yeast host, a useful promoter is the Saccharomyces cerevisiae enolase (eno-1) promoter. Particularly preferred promoters are the TAKA amylase, NA2-tpi (a hybrid of the promoters from the genes encoding Aspergillus niger neutral alpha-amylase and Aspergillus oryzae triose phosphate isomerase) and glaA promoters.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a terminator sequence at its 3′ terminus. The terminator sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any terminator which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred terminators are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase, Aspergillus niger alpha-glucosidase and Saccharomyces cerevisiae enolase.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred leaders are obtained from the genes encoding Aspergillus oryzae TAKA amylase and Aspergillus oryzae triose phosphate isomerase.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the fungal host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention, but particularly preferred polyadenylation sequences are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase and Aspergillus niger alpha-glucosidase.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed protein or fragment thereof within the cell, it is preferred that expression of the protein or fragment thereof gives rise to a product secreted outside the cell. To this end, a protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the fungal host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof. Alternatively, the 5′ end of the coding sequence may contain a signal peptide coding region which is foreign to that portion of the coding sequence which encodes the secreted protein or fragment thereof. The foreign signal peptide may be required where the coding sequence does not normally contain a signal peptide coding region. Alternatively, the foreign signal peptide may simply replace the natural signal peptide to obtain enhanced secretion of the desired protein or fragment thereof. The foreign signal peptide coding region may be obtained from a glucoamylase or an amylase gene from an Aspergillus species, a lipase or proteinase gene from Rhizomucor miehei, the gene for the alpha-factor from Saccharomyces cerevisiae, or the calf preprochymosin gene. An effective signal peptide for fungal host cells is the Aspergillus oryzae TAKA amylase signal, Aspergillus niger neutral amylase signal, the Rhizomucor miehei aspartic proteinase signal, the Humicola lanuginosus cellulase signal, or the Rhizomucor miehei lipase signal. However, any signal peptide capable of permitting secretion of the protein or fragment thereof in a fungal host of choice may be used in the present invention.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be linked to a propeptide coding region. A propeptide is an amino acid sequence found at the amino terminus of a proprotein or proenzyme. Cleavage of the propeptide from the proprotein yields a mature biochemically active protein. The resulting polypeptide is known as a propolypeptide or proenzyme (or a zymogen in some cases). Propolypeptides are generally inactive and can be converted to mature active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide or proenzyme. The propeptide coding region may be native to the protein or fragment thereof or may be obtained from foreign sources. The foreign propeptide coding region may be obtained from the Saccharomyces cerevisiae alpha-factor gene or Myceliophthora thermophila laccase gene (WO 95/33836, the entirety of which is herein incorporated by reference).

The procedures used to ligate the elements described above to construct the recombinant expression vector of the present invention are well known to one skilled in the art (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor, N.Y., (1989)).

The present invention also relates to recombinant fungal host cells produced by the methods of the present invention which are advantageously used with the recombinant vector of the present invention. The cell is preferably transformed with a vector comprising a nucleic acid sequence of the invention followed by integration of the vector into the host chromosome. The choice of fungal host cells will to a large extent depend upon the gene encoding the protein or fragment thereof and its source. The fungal host cell may, for example, be a yeast cell or afilamentous fungal cell.

“Yeast” as used herein includes Ascosporogenous yeast (Endomycetales), Basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti (Blastomycetes). The Ascosporogenous yeasts are divided into the families Spermophthoraceae and Saccharomycetaceae. The latter is comprised of four subfamilies, Schizosaccharomycoideae for example, genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae and Saccharomycoideae (for example, genera Pichia, Kluyveromyces and Saccharomyces). The Basidiosporogenous yeasts include the genera Leucosporidim, Rhodosporidium, Sporidiobolus, Filobasidium and Filobasidiella. Yeast belonging to the Fungi Imperfecti are divided into two families, Sporobolomycetaceae (for example, genera Sorobolomyces and Bullera) and Cryptococcaceae (for example, genus Candida). Since the classification of yeast may change in the future, for the purposes of this invention, yeast shall be defined as described in Biology and Activities of Yeast (Skinner et al., Soc. App. Bacteriol. Symposium Series No. 9, (1980), the entirety of which is herein incorporated by reference). The biology of yeast and manipulation of yeast genetics are well known in the art (see, for example, Biochemistry and Genetics of Yeast, Bacil et al. (ed.), 2nd edition, 1987; The Yeasts, Rose and Harrison (eds.), 2nd ed., (1987); and The Molecular Biology of the Yeast Saccharomyces , Strathern et al. (eds.), (1981), all of which are herein incorporated by reference in their entirety).

“Fungi” as used herein includes the phyla Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK; the entirety of which is herein incorporated by reference) as well as the Oomycota (as cited in Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK) and all mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). Representative groups of Ascomycota include, for example, Neurospora, Eupenicillium (=Penicillium), Emericella (=Aspergillus), Eurotiun (=Aspergillus) and the true yeasts listed above. Examples of Basidiomycota include mushrooms, rusts and smuts. Representative groups of Chytridiomycota include, for example, Allomyces, Blastocladiella, Coelomomyces and aquatic fungi. Representative groups of Oomycota include, for example, Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examples of mitosporic fungi include Aspergillus, Penicilliun, Candida and Alternaria. Representative groups of Zygomycota include, for example, Rhizopus and Mucor.

“Filamentous fungi” include all filamentous forms of the subdivision Eumycota and Oomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). The filamentous fungi are characterized by a vegetative mycelium composed of chitin, cellulose, glucan, chitosan, mannan and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. In contrast, vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of a unicellular thallus and carbon catabolism may be fermentative.

In one embodiment, the fungal host cell is a yeast cell. In a preferred embodiment, the yeast host cell is a cell of the species of Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia and Yarrowia. In a preferred embodiment, the yeast host cell is a Saccharomyces cerevisiae cell, a Saccharomyces carlsbergensis, Saccharomyces diastaticus cell, a Saccharomyces douglasii cell, a Saccharomyces kluyveri cell, a Saccharomyces norbensis cell, or a Saccharomyces oviformis cell. In another preferred embodiment, the yeast host cell is a Kluyveromyces lactis cell. In another preferred embodiment, the yeast host cell is a Yarrowia lipolytica cell.

In another embodiment, the fungal host cell is a filamentous fungal cell. In a preferred embodiment, the filamentous fungal host cell is a cell of the species of, but not limited to, Acremonium, Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium, Thielavia, Tolypocladium and Trichoderma. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus cell. In another preferred embodiment, the filamentous fungal host cell is an Acremonium cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora cell. In another even preferred embodiment, the filamentous fungal host cell is a Mucor cell. In another preferred embodiment, the filamentous fungal host cell is a Neurospora cell. In another preferred embodiment, the filamentous fungal host cell is a Penicillium cell. In another preferred embodiment, the filamentous fungal host cell is a Thielavia cell. In another preferred embodiment, the filamentous fungal host cell is a Tolypocladiun cell. In another preferred embodiment, the filamentous fungal host cell is a Trichoderma cell. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus oryzae cell, an Aspergillus niger cell, an Aspergillus foetidus cell, or an Aspergillus japonicus cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium oxysporum cell or a Fusarium graminearum cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola insolens cell or a Humicola lanuginosus cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora thermophila cell. In a most preferred embodiment, the filamentous fungal host cell is a Mucor miehei cell. In a most preferred embodiment, the filamentous fungal host cell is a Neurospora crassa cell. In a most preferred embodiment, the filamentous fungal host cell is a Penicillium purpurogenum cell. In another most preferred embodiment, the filamentous fungal host cell is a Thielavia terrestris cell. In another most preferred embodiment, the Trichoderma cell is a Trichoderma reesei cell, a Trichoderna viride cell, a Trichoderma longibrachiatum cell, a Trichoderma harzianum cell, or a Trichoderma koningii cell. In a preferred embodiment, the fungal host cell is selected from an A. nidulans cell, an A. niger cell, an A. oryzae cell and an A. sojae cell. In a further preferred embodiment, the fungal host cell is an A. nidulans cell.

The recombinant fungal host cells of the present invention may further comprise one or more sequences which encode one or more factors that are advantageous in the expression of the protein or fragment thereof, for example, an activator (e.g., a trans-acting factor), a chaperone and a processing protease. The nucleic acids encoding one or more of these factors are preferably not operably linked to the nucleic acid encoding the protein or fragment thereof. An activator is a protein which activates transcription of a nucleic acid sequence encoding a polypeptide (Kudla et al., EMBO 9:1355-1364(1990); Jarai and Buxton, Current Genetics 26:2238-244(1994); Verdier, Yeast 6:271-297(1990), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding an activator may be obtained from the genes encoding Saccharomyces cerevisiae heme activator protein 1 (hap1), Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4) and Aspergillus nidulans ammonia regulation protein (areA). For further examples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et al., Journal of Gen. Microbiol. 139:2295-2307 (1993), both of which are herein incorporated by reference in their entirety). A chaperone is a protein which assists another protein in folding properly (Hartl et al., TIBS 19:20-25 (1994); Bergeron et al., TIBS 19:124-128 (1994); Demolder et al., J. Biotechnology 32:179-189 (1994); Craig, Science 260:1902-1903(1993); Gething and Sambrook, Nature 355:33-45 (1992); Puig and Gilbert, J Biol. Chem. 269:7764-7771 (1994); Wang and Tsou, FASEB Journal 7:1515-11157 (1993); Robinson et al., Bio/Technology 1:381-384 (1994), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding a chaperone may be obtained from the genes encoding Aspergillus oryzae protein disulphide isomerase, Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae BiP/GRP78 and Saccharomyces cerevisiae sp70. For further examples, see Gething and Sambrook, Nature 355:33-45 (1992); Hartl et al., TIBS 19:20-25 (1994). A processing protease is a protease that cleaves a propeptide to generate a mature biochemically active polypeptide (Enderlin and Ogrydziak, Yeast 10:67-79 (1994); Fuller et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1434-1438 (1989); Julius et al., Cell 37:1075-1089 (1984); Julius et al., Cell 32:839-852 (1983), all of which are incorporated by reference in their entirety). The nucleic acid sequence encoding a processing protease may be obtained from the genes encoding Aspergillus niger Kex2, Saccharomyces cerevisiae dipeptidylaminopeptidase, Saccharomyces cerevisiae Kex2 and Yarrowia lipolytica dibasic processing endoprotease (xpr6). Any factor that is functional in the fungal host cell of choice may be used in the present invention.

Fungal cells may be transformed by a process involving protoplast formation, transformation of the protoplasts and regeneration of the cell wall in a manner known per se. Suitable procedures for transformation of Aspergillus host cells are described in EP 238 023 and Yelton et al., Proc. Natl. Acad. Sci. (U.S.A.) 81:1470-1474 (1984), both of which are herein incorporated by reference in their entirety. A suitable method of transforming Fusarium species is described by Malardier et al., Gene 78:147-156 (1989), the entirety of which is herein incorporated by reference. Yeast may be transformed using the procedures described by Becker and Guarente, In: Abelson and Simon, (eds.), Guide to Yeast Genetics and Molecular Biology, Methods Enzymol. Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., J. Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:1920 (1978), all of which are herein incorporated by reference in their entirety.

The present invention also relates to methods of producing the protein or fragment thereof comprising culturing the recombinant fungal host cells under conditions conducive for expression of the protein or fragment thereof. The fungal cells of the present invention are cultivated in a nutrient medium suitable for production of the protein or fragment thereof using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the protein or fragment thereof to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g., Bennett and LaSure (eds.), More Gene Manipulations in Fungi, Academic Press, CA, (1991), the entirety of which is herein incorporated by reference). Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection, Manassas, Va.). If the protein or fragment thereof is secreted into the nutrient medium, a protein or fragment thereof can be recovered directly from the medium. If the protein or fragment thereof is not secreted, it is recovered from cell lysates.

The expressed protein or fragment thereof may be detected using methods known in the art that are specific for the particular protein or fragment. These detection methods may include the use of specific antibodies, formation of an enzyme product, or disappearance of an enzyme substrate. For example, if the protein or fragment thereof has enzymatic activity, an enzyme assay may be used. Alternatively, if polyclonal or monoclonal antibodies specific to the protein or fragment thereof are available, immunoassays may be employed using the antibodies to the protein or fragment thereof. The techniques of enzyme assay and immunoassay are well known to those skilled in the art.

The resulting protein or fragment thereof may be recovered by methods known in the arts. For example, the protein or fragment thereof may be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The recovered protein or fragment thereof may then be further purified by a variety of chromatographic procedures, e.g., ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the like.

(c) Mammalian Constructs and Transformed Mammalian Cells

The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian host cell exogenous genetic material. The present invention also relates to a mammalian cell comprising a mammalian recombinant vector. The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC, Manassas, Va.), such as HeLa cells, Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells and a number of other cell lines. Suitable promoters for mammalian cells are also known in the art and include viral promoters such as that from Simian Virus 40 (SV40) (Fiers et al., Nature 273:113 (1978), the entirety of which is herein incorporated by reference), Rous sarcoma virus (RSV), adenovirus (ADV) and bovine papilloma virus (BPV). Mammalian cells may also require terminator sequences and poly-A addition sequences. Enhancer sequences which increase expression may also be included and sequences which promote amplification of the gene may also be desirable (for example methotrexate resistance genes).

Vectors suitable for replication in mammalian cells may include viral replicons, or sequences which insure integration of the appropriate sequences encoding HCV epitopes into the host genome. For example, another vector used to express foreign DNA is vaccinia virus. In this case, for example, a nucleic acid molecule encoding a protein or fragment thereof is inserted into the vaccinia genome. Techniques for the insertion of foreign DNA into the vaccinia virus genome are known in the art and may utilize, for example, homologous recombination. Such heterologous DNA is generally inserted into a gene which is non-essential to the virus, for example, the thymidine kinase gene (tk), which also provides a selectable marker. Plasmid vectors that greatly facilitate the construction of recombinant viruses have been described (see, for example, Mackett et al, J Virol. 49:857 (1984); Chakrabarti et al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer Vectors For Mammalian Cells (Miller and Calos, eds., Cold Spring Harbor Laboratory, N.Y., p. 10, (1987); all of which are herein incorporated by reference in their entirety). Expression of the HCV polypeptide then occurs in cells or animals which are infected with the live recombinant vaccinia virus.

The sequence to be integrated into the mammalian sequence may be introduced into the primary host by any convenient means, which includes calcium precipitated DNA, spheroplast fusion, transformation, electroporation, biolistics, lipofection, microinjection, or other convenient means. Where an amplifiable gene is being employed, the amplifiable gene may serve as the selection marker for selecting hosts into which the amplifiable gene has been introduced. Alternatively, one may include with the amplifiable gene another marker, such as a drug resistance marker, e.g. neomycin resistance (G418 in mammalian cells), hygromycin in resistance etc., or an auxotrophy marker (HIS3, TRP1, LEU2, URA3, ADE2, LYS2, etc.) for use in yeast cells.

Depending upon the nature of the modification and associated targeting construct, various techniques may be employed for identifying targeted integration. Conveniently, the DNA may be digested with one or more restriction enzymes and the fragments probed with an appropriate DNA fragment which will identify the properly sized restriction fragment associated with integration.

One may use different promoter sequences, enhancer sequences, or other sequence which will allow for enhanced levels of expression in the expression host. Thus, one may combine an enhancer from one source, a promoter region from another source, a 5′-noncoding region upstream from the initiation sucrose from the same or different source as the other sequences and the like. One may provide for an intron in the non-coding region with appropriate splice sites or for an alternative 3′-untranslated sequence or polyadenylation site. Depending upon the particular purpose of the modification, any of these sequences may be introduced, as desired.

Where selection is intended, the sequence to be integrated will have with it a marker gene, which allows for selection. The marker gene may conveniently be downstream from the target gene and may include resistance to a cytotoxic agent, e.g. antibiotics, heavy metals, or the like, resistance or susceptibility to HAT, gancyclovir, etc., complementation to an auxotrophic host, particularly by using an auxotrophic yeast as the host for the subject manipulations, or the like. The marker gene may also be on a separate DNA molecule, particularly with primary mammalian cells. Alternatively, one may screen the various transformants, due to the high efficiency of recombination in yeast, by using hybridization analysis, PCR, sequencing, or the like.

For homologous recombination, constructs can be prepared where the amplifiable gene will be flanked, normally on both sides with DNA homologous with the DNA of the target region. Depending upon the nature of the integrating DNA and the purpose of the integration, the homologous DNA will generally be within 100 kb, usually 50 kb, preferably about 25 kb, of the transcribed region of the target gene, more preferably within 2 kb of the target gene. Where modeling of the gene is intended, homology will usually be present proximal to the site of the mutation. The homologous DNA may include the 5′-upstream region outside of the transcriptional regulatory region or comprising any enhancer sequences, transcriptional initiation sequences, adjacent sequences, or the like. The homologous region may include a portion of the coding region, where the coding region may be comprised only of an open reading frame or combination of exons and introns. The homologous region may comprise all or a portion of an intron, where all or a portion of one or more exons may also be present. Alternatively, the homologous region may comprise the 3′-region, so as to comprise all or a portion of the transcriptional termination region, or the region 3′ of this region. The homologous regions may extend over all or a portion of the target gene or be outside the target gene comprising all or a portion of the transcriptional regulatory regions and/or the structural gene.

The integrating constructs may be prepared in accordance with conventional ways, where sequences may be synthesized, isolated from natural sources, manipulated, cloned, ligated, subjected to in vitro mutagenesis, primer repair, or the like. At various stages, the joined sequences may be cloned and analyzed by restriction analysis, sequencing, or the like. Usually during the preparation of a construct where various fragments are joined, the fragments, intermediate constructs and constructs will be carried on a cloning vector comprising a replication system functional in a prokaryotic host, e.g., E. coli and a marker for selection, e.g., biocide resistance, complementation to an auxotrophic host, etc. Other functional sequences may also be present, such as polylinkers, for ease of introduction and excision of the construct or portions thereof, or the like. A large number of cloning vectors are available such as pBR322, the pUC series, etc. These constructs may then be used for integration into the primary mammalian host.

In the case of the primary mammalian host, a replicating vector may be used. Usually, such vector will have a viral replication system, such as SV40, bovine papilloma virus, adenovirus, or the like. The linear DNA sequence vector may also have a selectable marker for identifying transfected cells. Selectable markers include the neo gene, allowing for selection with G418, the herpes tk gene for selection with HAT medium, the gpt gene with mycophenolic acid, complementation of an auxotrophic host, etc.

The vector may or may not be capable of stable maintenance in the host. Where the vector is capable of stable maintenance, the cells will be screened for homologous integration of the vector into the genome of the host, where various techniques for curing the cells may be employed. Where the vector is not capable of stable maintenance, for example, where a temperature sensitive replication system is employed, one may change the temperature from the permissive temperature to the non-permissive temperature, so that the cells may be cured of the vector. In this case, only those cells having integration of the construct comprising the amplifiable gene and, when present, the selectable marker, will be able to survive selection.

Where a selectable marker is present, one may select for the presence of the targeting construct by means of the selectable marker. Where the selectable marker is not present, one may select for the presence of the construct by the amplifiable gene. For the neo gene or the herpes tk gene, one could employ a medium for growth of the transformants of about 0. 1-1 mg/ml of G418 or may use HAT medium, respectively. Where DHFR is the amplifiable gene, the selective medium may include from about 0.01-0.5 M of methotrexate or be deficient in glycine-hypoxanthine-thymidine and have dialysed serum (GHT media).

The DNA can be introduced into the expression host by a variety of techniques that include calcium phosphate/DNA co-precipitates, microinjection of DNA into the nucleus, electroporation, yeast protoplast fusion with intact cells, transfection, polycations, e.g., polybrene, polyornithine, etc., or the like. The DNA may be single or double stranded DNA, linear or circular. The various techniques for transforming mammalian cells are well known (see Keown et al., Methods Enzymol. (1989); Keown et al., Methods Enzymol. 185:527-537 (1990); Mansour et al., Nature 336:348-352, (1988); all of which are herein incorporated by reference in their entirety).

(d) Insect Constructs and Transformed Insect Cells

The present invention also relates to an insect recombinant vectors comprising exogenous genetic material. The present invention also relates to an insect cell comprising an insect recombinant vector. The present invention also relates to methods for obtaining a recombinant insect host cell, comprising introducing into an insect cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

The insect recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of the nucleic acid sequence. The choice of a vector will typically depend on the compatibility of the vector with the insect host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the insect host. In addition, the insect vector may be an expression vector. Nucleic acid molecules can be suitably inserted into a replication vector for expression in the insect cell under a suitable promoter for insect cells. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid molecule to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for insect cell transformation generally include, but are not limited to, one or more of the following: a signal sequence, origin of replication, one or more marker genes and an inducible promoter.

The insect vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the insect cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the insect host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the insect host cell and, furthermore, may be non-encoding or encoding sequences.

Baculovirus expression vectors (BEVs) have become important tools for the expression of foreign genes, both for basic research and for the production of proteins with direct clinical applications in human and veterinary medicine (Doerfler, Curr. Top. Microbiol. Immunol. 131:51-68 (1968); Luckow and Summers, Bio/Technology 6:47-55 (1988a); Miller, Annual Review of Microbiol. 42:177-199 (1988); Summers, Curr. Comm. Molecular Biology, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988); all of which are herein incorporated by reference in their entirety). BEVs are recombinant insect viruses in which the coding sequence for a chosen foreign gene has been inserted behind a baculovirus promoter in place of the viral gene, e.g., polyhedrin (Smith and Summers, U.S. Pat. No. 4,745,051, the entirety of which is incorporated herein by reference).

The use of baculovirus vectors relies upon the host cells being derived from Lepidopteran insects such as Spodoptera frugiperda or Trichoplusia ni. The preferred Spodoptera frugiperda cell line is the cell line Sf9. The Spodoptera frugiperda Sf9 cell line was obtained from American Type Culture Collection (Manassas, Va.) and is assigned accession number ATCC CRL 1711 (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), the entirety of which is herein incorporated by reference). Other insect cell systems, such as the silkworm B. mori may also be used.

The proteins expressed by the BEVs are, therefore, synthesized, modified and transported in host cells derived from Lepidopteran insects. Most of the genes that have been inserted and produced in the baculovirus expression vector system have been derived from vertebrate species. Other baculovirus genes in addition to the polyhedrin promoter may be employed to advantage in a baculovirus expression system. These include immediate-early (alpha), delayed-early ( ), late ( ), or very late (delta), according to the phase of the viral infection during which they are expressed. The expression of these genes occurs sequentially, probably as the result of a “cascade” mechanism of transcriptional regulation. (Guarino and Summers, J. Virol. 57:563-571 (1986); Guarino and Summers, J. Virol. 61:2091-2099 (1987); Guarino and Summers, Virol. 162:444-451 (1988); all of which are herein incorporated by reference in their entirety).

Insect recombinant vectors are useful as intermediates for the infection or transformation of insect cell systems. For example, an insect recombinant vector containing a nucleic acid molecule encoding a baculovirus transcriptional promoter followed downstream by an insect signal DNA sequence is capable of directing the secretion of the desired biologically active protein from the insect cell. The vector may utilize a baculovirus transcriptional promoter region derived from any of the over 500 baculoviruses generally infecting insects, such as for example the Orders Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera, including for example but not limited to the viral DNAs of Autographa californica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV, Rachiplusia ou MNPV or Galleria mellonella MNPV, wherein said baculovirus transcriptional promoter is a baculovirus immediate-early gene IE1 or IEN promoter; an immediate-early gene in combination with a baculovirus delayed-early gene promoter region selected from the group consisting of 39K and a HindIII-k fragment delayed-early gene; or a baculovirus late gene promoter. The immediate-early or delayed-early promoters can be enhanced with transcriptional enhancer elements. The insect signal DNA sequence may code for a signal peptide of a Lepidopteran adipokinetic hormone precursor or a signal peptide of the Manduca sexta adipokinetic hormone precursor (Summers, U.S. Pat. No. 5,155,037; the entirety of which is herein incorporated by reference). Other insect signal DNA sequences include a signal peptide of the Orthoptera Schistocerca gregaria locust adipokinetic hormone precurser and the Drosophila melanogaster cuticle genes CP1, CP2, CP3 or CP4 or for an insect signal peptide having substantially a similar chemical composition and function (Summers, U.S. Pat. No. 5,155,037).

Insect cells are distinctly different from animal cells. Insects have a unique life cycle and have distinct cellular properties such as the lack of intracellular plasminogen activators in which are present in vertebrate cells. Another difference is the high expression levels of protein products ranging from 1 to greater than 500 mg/liter and the ease at which cDNA can be cloned into cells (Frasier, In Vitro Cell. Dev. Biol. 25:225 (1989); Summers and Smith, In: A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), both of which are incorporated by reference in their entirety).

Recombinant protein expression in insect cells is achieved by viral infection or stable transformation. For viral infection, the desired gene is cloned into baculovirus at the site of the wild-type polyhedron gene (Webb and Summers, Technique 2:173 (1990); Bishop and Posse, Adv. Gene Technol. 1:55 (1990); both of which are incorporated by reference in their entirety). The polyhedron gene is a component of a protein coat in occlusions which encapsulate virus particles. Deletion or insertion in the polyhedron gene results the failure to form occlusion bodies. Occlusion negative viruses are morphologically different from occlusion positive viruses and enable one skilled in the art to identify and purify recombinant viruses.

The vectors of present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. Selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, a nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the insect host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof. The promoter may be any nucleic acid sequence which shows transcriptional activity in the insect host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell.

For example, a nucleic acid molecule encoding a protein or fragment thereof may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the insect host cell of choice may be used in the present invention.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the insect host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed polypeptide within the cell, it is preferred that expression of the polypeptide gene gives rise to a product secreted outside the cell. To this end, the protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the insect host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof.

At present, a mode of achieving secretion of a foreign gene product in insect cells is by way of the foreign gene's native signal peptide. Because the foreign genes are usually from non-insect organisms, their signal sequences may be poorly recognized by insect cells and hence, levels of expression may be suboptimal. However, the efficiency of expression of foreign gene products seems to depend primarily on the characteristics of the foreign protein. On average, nuclear localized or non-structural proteins are most highly expressed, secreted proteins are intermediate and integral membrane proteins are the least expressed. One factor generally affecting the efficiency of the production of foreign gene products in a heterologous host system is the presence of native signal sequences (also termed presequences, targeting signals, or leader sequences) associated with the foreign gene. The signal sequence is generally coded by a DNA sequence immediately following (5′ to 3′) the translation start site of the desired foreign gene.

The expression dependence on the type of signal sequence associated with a gene product can be represented by the following example: If a foreign gene is inserted at a site downstream from the translational start site of the baculovirus polyhedrin gene so as to produce a fusion protein (containing the N-terminus of the polyhedrin structural gene), the fused gene is highly expressed. But less expression is achieved when a foreign gene is inserted in a baculovirus expression vector immediately following the transcriptional start site and totally replacing the polyhedrin structural gene.

Insertions into the region −50 to −1 significantly alter (reduce) steady state transcription which, in turn, reduces translation of the foreign gene product. Use of the pVL941 vector optimizes transcription of foreign genes to the level of the polyhedrin gene transcription. Even though the transcription of a foreign gene may be optimal, optimal translation may vary because of several factors involving processing: signal peptide recognition, mRNA and ribosome binding, glycosylation, disulfide bond formation, sugar processing, oligomerization, for example.

The properties of the insect signal peptide are expected to be more optimal for the efficiency of the translation process in insect cells than those from vertebrate proteins. This phenomenon can generally be explained by the fact that proteins secreted from cells are synthesized as precursor molecules containing hydrophobic N-terminal signal peptides. The signal peptides direct transport of the select protein to its target membrane and are then cleaved by a peptidase on the membrane, such as the endoplasmic reticulum, when the protein passes through it.

Another exemplary insect signal sequence is the sequence encoding for Drosophila cuticle proteins such as CP1, CP2, CP3 or CP4 (Summers, U.S. Pat. No. 5,278,050; the entirety of which is herein incorporated by reference). Most of a 9 kb region of the Drosophila genome containing genes for the cuticle proteins has been sequenced. Four of the five cuticle genes contains a signal peptide coding sequence interrupted by a short intervening sequence (about 60 base pairs) at a conserved site. Conserved sequences occur in the 5′ mRNA untranslated region, in the adjacent 35 base pairs of upstream flanking sequence and at −200 base pairs from the mRNA start position in each of the cuticle genes.

Standard methods of insect cell culture, cotransfection and preparation of plasmids are set forth in Summers and Smith (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Agricultural Experiment Station Bulletin No. 1555, Texas A&M University (1987)). Procedures for the cultivation of viruses and cells are described in Volkman and Summers, J. Virol 19:820-832 (1975) and Volkman et al., J. Virol 19:820-832 (1976); both of which are herein incorporated by reference in their entirety.

(e) Bacterial Constructs and Transformed Bacterial Cells

The present invention also relates to a bacterial recombinant vector comprising exogenous genetic material. The present invention also relates to a bacteria cell comprising a bacterial recombinant vector. The present invention also relates to methods for obtaining a recombinant bacteria host cell, comprising introducing into a bacterial host cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

The bacterial recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the bacterial host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the bacterial host. In addition, the bacterial vector may be an expression vector. Nucleic acid molecules encoding protein homologues or fragments thereof can, for example, be suitably inserted into a replicable vector for expression in the bacterium under the control of a suitable promoter for bacteria. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for bacterial transformation generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes and an inducible promoter.

In general, plasmid vectors containing replicon and control sequences that are derived from species compatible with the host cell are used in connection with bacterial hosts. The vector ordinarily carries a replication site, as well as marking sequences that are capable of providing phenotypic selection in transformed cells. For example, E. coli is typically transformed using pBR322, a plasmid derived from an E. coli species (see, e.g., Bolivar et al., Gene 2:95 (1977); the entirety of which is herein incorporated by reference). pBR322 contains genes for ampicillin and tetracycline resistance and thus provides easy means for identifying transformed cells. The pBR322 plasmid, or other microbial plasmid or phage, also generally contains, or is modified to contain, promoters that can be used by the microbial organism for expression of the selectable marker genes.

Nucleic acid molecules encoding protein or fragments thereof may be expressed not only directly, but also as a fusion with another polypeptide, preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature polypeptide. In general, the signal sequence may be a component of the vector, or it may be a part of the polypeptide DNA that is inserted into the vector. The heterologous signal sequence selected should be one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For bacterial host cells that do not recognize and process the native polypeptide signal sequence, the signal sequence is substituted by a bacterial signal sequence selected, for example, from the group consisting of the alkaline phosphatase, penicillinase, lpp, or heat-stable enterotoxin II leaders.

Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA and, includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria.

Expression and cloning vectors also generally contain a selection gene, also termed a selectable marker. This gene encodes a protein necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene will not survive in the culture medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous protein homologue or fragment thereof produce a protein conferring drug resistance and thus survive the selection regimen.

The expression vector for producing a protein or fragment thereof can also contains an inducible promoter that is recognized by the host bacterial organism and is operably linked to the nucleic acid encoding, for example, the nucleic acid molecule encoding the protein homologue or fragment thereof of interest. Inducible promoters suitable for use with bacterial hosts include the -lactamase and lactose promoter systems (Chang et al., Nature 275:615 (1978); Goeddel et al., Nature 281:544 (1979); both of which are herein incorporated by reference in their entirety), the arabinose promoter system (Guzman et al., J. Bacteriol. 174:7716-7728 (1992); the entirety of which is herein incorporated by reference), alkaline phosphatase, a tryptophan (trp) promoter system (Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both of which are herein incorporated by reference in their entirety) and hybrid promoters such as the tac promoter (deBoer et al., Proc. Natl. Acad. Sci. (USA) 80:21-25 (1983); the entirety of which is herein incorporated by reference). However, other known bacterial inducible promoters are suitable (Siebenlist et al., Cell 20:269 (1980); the entirety of which is herein incorporated by reference).

Promoters for use in bacterial systems also generally contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide of interest. The promoter can be removed from the bacterial source DNA by restriction enzyme digestion and inserted into the vector containing the desired DNA.

Construction of suitable vectors containing one or more of the above-listed components employs standard ligation techniques. Isolated plasmids or DNA fragments are cleaved, tailored and re-ligated in the form desired to generate the plasmids required. Examples of available bacterial expression vectors include, but are not limited to, the multifunctional E. coli cloning and expression vectors such as Bluescript™ (Stratagene, La Jolla, Calif.), in which, for example, encoding an A. nidulans protein homologue or fragment thereof homologue, may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of -galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke and Schuster, J. Biol. Chem. 264:5503-5509 (1989), the entirety of which is herein incorporated by reference); and the like. pGEX vectors (Promega, Madison, Wis. U.S.A.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems are designed to include heparin, thrombin or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.

Suitable host bacteria for a bacterial vector include archaebacteria and eubacteria, especially eubacteria and most preferably Enterobacteriaceae. Examples of useful bacteria include Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla and Paracoccus. Suitable E. coli hosts include E. coli W3110 (American Type Culture Collection (ATCC) 27,325, Manassas, Va. U.S.A.), E. coli 294 (ATCC 31,446), E. coli B and E. coli X1776 (ATCC 31,537). These examples are illustrative rather than limiting. Mutant cells of any of the above-mentioned bacteria may also be employed. It is, of course, necessary to select the appropriate bacteria taking into consideration replicability of the replicon in the cells of a bacterium. For example, E. coli, Serratia, or Salmonella species can be suitably used as the host when well known plasmids such as pBR322, pBR325, pACYC177, or pKN410 are used to supply the replicon. E. coli strain W3110 is a preferred host or parent host because it is a common host strain for recombinant DNA product fermentations. Preferably, the host cell should secrete minimal amounts of proteolytic enzymes.

Host cells are transfected and preferably transformed with the above-described vectors and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.

Numerous methods of transfection are known to the ordinarily skilled artisan, for example, calcium phosphate and electroporation. Depending on the host cell used, transformation is done using standard techniques appropriate to such cells. The calcium treatment employing calcium chloride, as described in section 1.82 of Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989), is generally used for bacterial cells that contain substantial cell-wall barriers. Another method for transformation employs polyethylene glycol/DMSO, as described in Chung and Miller (Chung and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety of which is herein incorporated by reference). Yet another method is the use of the technique termed electroporation.

Bacterial cells used to produce the polypeptide of interest for purposes of this invention are cultured in suitable media in which the promoters for the nucleic acid encoding the heterologous polypeptide can be artificially induced as described generally, e.g., in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989). Examples of suitable media are given in U.S. Pat. Nos. 5,304,472 and 5,342,763; both of which are incorporated by reference in their entirety.

In addition to the above discussed procedures, practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of macromolecules (e.g., DNA molecules, plasmids, etc.), generation of recombinant organisms and the screening and isolating of clones, (see for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989); Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995), the entirety of which is herein incorporated by reference; Birren et al., Genome Analysis: Analyzing DNA, 1, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference).

(f) Computer Readable Media

The nucleotide sequence provided in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof, or complement thereof, or a nucleotide sequence at least 90% identical, preferably 95%, identical even more preferably 99% or 100% identical to the sequence provided in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof, or complement thereof, can be “provided” in a variety of mediums to facilitate use. Such a medium can also provide a subset thereof in a form that allows a skilled artisan to examine the sequences.

A preferred subset of nucleotide sequences are those nucleic acid sequences that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragmnt of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof or fragment of either.

A further preferred subset of nucleic acid sequences is where the subset of sequences is two proteins or fragments thereof, more preferably three proteins or fragments thereof and even more preferable four proteins or fragments thereof, these nucleic acid sequences are selected from the group that comprises a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H⁺ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophophorylase enzyme or complement thereof or fragment of either.

In one application of this embodiment, a nucleotide sequence of the present invention can be recorded on computer readable media. As used herein, “computer readable media” refers to any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc, storage medium and magnetic tape: optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. A skilled artisan can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising computer readable medium having recorded thereon a nucleotide sequence of the present invention.

As used herein, “recorded” refers to a process for storing information on computer readable medium. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable medium to generate media comprising the nucleotide sequence information of the present invention. A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a nucleotide sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of data processor structuring formats (e.g. text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

By providing one or more of nucleotide sequences of the present invention, a skilled artisan can routinely access the sequence information for a variety of purposes. Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium. The examples which follow demonstrate how software which implements the BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990), the entirety of which is herein incorporated by reference) and BLAZE (Brutlag et al., Comp. Chem. 17:203-207 (1993), the entirety of which is herein incorporated by reference) search algorithms on a Sybase system can be used to identify open reading frames (ORFs) within the genome that contain homology to ORFs or proteins from other organisms. Such ORFs are protein-encoding fragments within the sequences of the present invention and are useful in producing commercially important proteins such as enzymes used in amino acid biosynthesis, metabolism, transcription, translation, RNA processing, nucleic acid and a protein degradation, protein modification and DNA replication, restriction, modification, recombination and repair.

The present invention further provides systems, particularly computer-based systems, which contain the sequence information described herein. Such systems are designed to identify commercially important fragments of the nucleic acid molecule of the present invention. As used herein, “a computer-based system” refers to the hardware means, software means and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based system are suitable for use in the present invention.

As indicated above, the computer-based systems of the present invention comprise a data storage means having stored therein a nucleotide sequence of the present invention and the necessary hardware means and software means for supporting and implementing a search means. As used herein, “data storage means” refers to memory that can store nucleotide sequence information of the present invention, or a memory access means which can access manufactures having recorded thereon the nucleotide sequence information of the present invention. As used herein, “search means” refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence information stored within the data storage means. Search means are used to identify fragments or regions of the sequence of the present invention that match a particular target sequence or target motif. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are available can be used in the computer-based systems of the present invention. Examples of such software include, but are not limited to, MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of the available algorithms or implementing software packages for conducting homology searches can be adapted for use in the present computer-based systems.

The most preferred sequence length of a target sequence is from about 10 to 100 amino acids or from about 30 to 300 nucleotide residues. However, it is well recognized that during searches for commercially important fragments of the nucleic acid molecules of the present invention, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

As used herein, “a target structural motif,” or “target motif,” refers to any rationally selected sequence or combination of sequences in which the sequences the sequence(s) are chosen based on a three-dimensional configuration which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzymatic active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, promoter sequences, cis elements, hairpin structures and inducible expression elements (protein binding sequences).

Thus, the present invention further provides an input means for receiving a target sequence, a data storage means for storing the target sequences of the present invention sequence identified using a search means as described above and an output means for outputting the identified homologous sequences. A variety of structural formats for the input and output means can be used to input and output information in the computer-based systems of the present invention. A preferred format for an output means ranks fragments of the sequence of the present invention by varying degrees of homology to the target sequence or target motif. Such presentation provides a skilled artisan with a ranking of sequences which contain various amounts of the target sequence or target motif and identifies the degree of homology contained in the identified fragment.

A variety of comparing means can be used to compare a target sequence or target motif with the data storage means to identify sequence fragments sequence of the present invention. For example, implementing software which implement the BLAST and BLAZE algorithms (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) can be used to identify open frames within the nucleic acid molecules of the present invention. A skilled artisan can readily recognize that any one of the publicly available homology search programs can be used as the search means for the computer-based systems of the present invention.

Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by way of illustration and are not intended to be limiting of the present invention, unless specified.

EXAMPLE 1

The MONN01 cDNA library is a normalized library generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON001 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) immature tassels at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON003 library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) roots at the V6 developmental stage. Seeds are planted at a depth of approximately 3 cm in coil into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, the seedlings are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting at a concentration of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in approximately 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6 leaf development stage. The root system is cut from maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON004 cDNA library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON005 cDNA library is generated from maize (B73xMo17, Illinois Foundation Seeds, Champaign, Ill., U.S.A.) root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON006 cDNA library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill., U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON007 cDNA library is generated from the primary root tissue of 5 day old maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). After germination, the trays, along with the moist paper, are moved to a greenhouse where the maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles for approximately 5 days. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. The primary root tissue is collected when the seedlings are 5 days old. At this stage, the primary root (radicle) is pushed through the coleorhiza which itself is pushed through the seed coat. The primary root, which is about 2-3 cm long, is cut and immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON008 cDNA library is generated from the primary shoot (coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings which are approximately 5 days old. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to a greenhouse at 15hr daytime/9 hr nighttime cycles and grown until they are 5 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 5 days old. At this stage, the primary shoot (coleoptile) is pushed through the seed coat and is about 2-3 cm long. The coleoptile is dissected away from the rest of the seedling, immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON009 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves at the 8 leaf stage (V8 plant development stage). Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80OF and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 8-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical, are cut at the base of the leaves. The leaves are then pooled and then immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON010 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the V8 development stage. The root system is cut from this mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON011 cDNA library is generated from undeveloped maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The second youngest leaf which is at the base of the apical leaf of V6 stage maize plant is cut at the base and immediately transferred to liquid nitrogen containers in which the leaf is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON012 cDNA library is generated from 2 day post germination maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to the greenhouse and grown at 15hr daytime/9 hr nighttime cycles until 2 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 2 days old. At the two day stage, the coleorhiza is pushed through the seed coat and the primary root (the radicle) is pierced the coleorhiza but is barely visible. Also, at this two day stage, the coleoptile is just emerging from the seed coat. The 2 days post germination seedlings are then immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° C. until preparation of total RNA.

The SATMON013 cDNA library is generated from apical maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) meristem founder at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, the plant is at the 4 leaf stage. The lead at the apex of the V4 stage maize plant is referred to as the meristem founder. This apical meristem founder is cut, immediately frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON014 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm fourteen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the maize plant ear shoots are ready for fertilization. At this stage, the ear shoots are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are pollinated and 14 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON016 library is a maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) sheath library collected at the V8 developmental stage. Seeds are planted in a depth of approximately 3 cm in solid into 2-3 inch pots containing Metro growing medium. After 2-3 weeks growth, they are transplanted into 10″ pots containing the same. Plants are watered daily before transplantation and approximately the times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plants are at the V8 stage the 5^(th) and 6^(th) leaves from the bottom exhibit fully developed leaf blades. At the base of these leaves, the ligule is differentiated and the leaf blade is joined to the sheath. The sheath is dissected away from the base of the leaf then the sheath is frozen in liquid nitrogen and crushed. The tissue is then stored at −80° C. until RNA preparation.

The SATMON017 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo seventeen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth the seeds are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are fertilized and 21 days after pollination, the ears are pulled out and the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON019 (Lib3054) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) culm (stem) at the V8 developmental stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plant is at the V8 stage, the 5th and 6th leaves from the bottom have fully developed leaf blades. The region between the nodes of the 5th and the sixth leaves from the bottom is the region of the stem that is collected. The leaves are pulled out and the sheath is also torn away from the stem. This stem tissue is completely free of any leaf and sheath tissue. The stem tissue is then frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The SATMON020 cDNA library is from a maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Initiated Callus. Petri plates containing approximately 25 ml of Type II initiation media are prepared. This medium contains N6 salts and vitamins, 3% sucrose, 2.3 g/liter proline 0.1 g/liter enzymatic casein hydrolysate, 2mg/liter 2,4-dichloro phenoxy-acetic acid (2,4, D), 15.3 mg/liter AgNO₃ and 0.8% bacto agar and is adjusted to pH 6.0 before autoclaving. At 9-11 days after pollination, an ear with immature embryos measuring approximately 1-2 mm in length is chosen. The husks and silks are removed and then the ear is broken into halves and placed in an autoclaved solution of Clorox/TWEEN 20 sterilizing solution. Then the ear is rinsed with deionized water. Then each embryo is extracted from the kernel. Intact embryos are placed in contact with the medium, scutellar side up). Multiple embryos are plated on each plate and the plates are incubated in the dark at 25° C. Type II calluses are friable, can be subcultured with a spatula, frequently regenerate via somatic embryogenesis and are relatively undifferentiated. As seen in the microscope, the Tape II calluses show color ranging from translucent to light yellow and heterogeneity on with respect to embryoid structure as well as stage of embryoid development. Once Type II callus are formed, the calluses is transferred to type II callus maintenance medium without AgNO₃. Every 7-10 days, the callus is subcultured. About 4 weeks after embryo isolation the callus is removed from the plates and then frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON021 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. As the maize plant enters the V8 stage, tassels which are 15-20 cm in length are collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON022 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silks) at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the plant is in the V8 stage. At this stage, some immature ear shoots are visible. The immature ear shoots (approximately 1 cm in length) are pulled out, frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON23 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) ear (growing silk) at the V8 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. When the tissue is harvested at the V8 stage, the length of the ear that is harvested is about 10-15 cm and the silks are just exposed (approximately 1 inch). The ear along with the silks is frozen in liquid nitrogen and then the tissue is stored at −80° C. until RNA preparation.

The SATMON024 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the V9 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. As a maize plant enters the V9 stage, the tassel is rapidly developing and a 37 cm tassel along with the glume, anthers and pollen is collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON025 cDNA library is from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) Hill Type II-Regenerated Callus. Type II callus is grown in initiation media as described for SATMON020 and then the embryoids on the surface of the Type II callus are allowed to mature and germinate. The 1-2 gm fresh weight of the soft friable type callus containing numerous embryoids are transferred to 100×15 mm petri plates containing 25 ml of regeneration media. Regeneration media consists of Murashige and Skoog (MS) basal salts, modified White's vitamins (0.2 g/liter glycine and 0.5 g/liter myo-inositoland 0.8% bacto agar (6SMS0D)). The plates are then placed in the dark after covering with parafilm. After 1 week, the plates are moved to a lighted growth chamber with 16 hr light and 8 hr dark photoperiod. Three weeks after plating the Type II callus to 6SMS0D, the callus exhibit shoot formation. The callus and the shoots are transferred to fresh 6SMS0D plates for another 2 weeks. The callus and the shoots are then transferred to petri plates with reduced sucrose (3SMS0D). Upon distinct formation of a root and shoot, the newly developed green plants are then removed out with a spatula and frozen in liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON026 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) juvenile/adult shift leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plants are at the 8-leaf development stage. Leaves are founded sequentially around the meristem over weeks of time and the older, more juvenile leaves arise earlier and in a more basal position than the younger, more adult leaves, which are in a more apical position. In a V8 plant, some leaves which are in the middle portion of the plant exhibit characteristics of both juvenile as well as adult leaves. They exhibit a yellowing color but also exhibit, in part, a green color. These leaves are termed juvenile/adult shift leaves. The juvenile/adult shift leaves (the 4th, 5th leaves from the bottom) are cut at the base, pooled and transferred to liquid nitrogen in which they are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON027 cDNA library is generated from 6 day maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaves. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical, are all cut at the base of the leaves. All the leaves exhibit significant wilting. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON028 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) roots at the V8 developmental stage that are subject to six days water stress. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The root system is cut, shaken and washed to remove soil. Root tissue is then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON029 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings at the etiolated stage. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark for 4 days at approximately 70° F. Tissue is collected when the seedlings are 4 days old. By 4 days, the primary root has penetrated the coleorhiza and is about 4-5 cm and the secondary lateral roots have also made their appearance. The coleoptile has also pushed through the seed coat and is about 4-5 cm long. The seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON030 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) root tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, they are transplanted into 10 inch pots containing the same. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant, from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 1 5hr day/9hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 sodium vapor lamps. Tissue is collected when the maize plant is at the 4 leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON031 cDNA library is generated from the maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 4-leaf development stage. The third leaf from the bottom is cut at the base and immediately frozen in liquid nitrogen and crushed. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON033 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) embryo tissue 13 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 13 days after pollination, the ears are pulled out and then the kernels are plucked cut of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON034 cDNA library is generated from cold stressed maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept on at 10° C. for 7 days. After 7 days, the temperature is shifted to 15° C. for one day until germination of the seed. Tissue is collected once the seedlings are 1 day old. At this point, the coleorhiza has just pushed out of the seed coat and the primary root is just making its appearance. The coleoptile has not yet pushed completely through the seed coat and is also just making its appearance. These 1 day old cold stressed seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON˜001 (Lib36, Lib83, Lib84) cDNA library is generated from maize leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V8 stage. The older more juvenile leaves in a basal position was well as the younger more adult leaves which are more apical are all cut at the base, pooled and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMONN01 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) normalized immature tassels at the V6 plant development stage normalized tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN04 cDNA library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill. U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN05 cDNA library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill., U.S.A.) normalized root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation. The single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN06 cDNA library is generated from maize (B73×Mo17, Illinois Foundation Seeds, Champaign, Ill., U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated DATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The CMZ029 (SATMON036) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm 22 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 22 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the alurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The CMz030 (Lib143) cDNA library is generated from maize seedling tissue two days post germination. Seeds are planted on a moist filter paper on a covered try that is keep in the dark until germination. The trays are then moved to the bench top at 15 hr daytime/9 hr nighttime cycles for 2 days post-germination. The day time temperature is 80° F. and the nighttime temperature is 70° F. Tissue is collected when the seedlings are 2 days old. At this stage, the colehrhiza has pushed through the seed coat and the primary root (the radicle) is just piercing the colehrhiza and is barely visible. The seedlings are placed at 42° C. for 1 hour. Following the heat shock treatment, the seedlings are immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° until RNA preparation.

The CMz031 (Lib148) cDNA library is generated from maize pollen tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag to withhold pollen. Twenty-one days after pollination, prior to removing the ears, the paper bag is shaken to collect the mature pollen. The mature pollen is immediately frozen in liquid nitrogen containers and the pollen is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz033 (Lib189) cDNA library is generated from maize pooled leaf tissue. Samples are harvested from open pollinated plants. Tissue is collected from maize leaves at the anthesis stage. The leaves are collect from 10-12 plants and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz034 (Lib3060) cDNA library is generated from maize mature tissue at 40 days post pollination plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from leaves located two leaves below the ear leaf. This sample represents those genes expressed during onset and early stages of leaf senescence. The leaves are pooled and immediately transferred to liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz035 (Lib3061) cDNA library is generated from maize endosperm tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80 F and the nighttime temperature is approximately 70 F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence to withhold pollen. Thirty-two days after pollination, the ears are pulled out and the kernels are removed from the cob. Each kernel is dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately transferred to liquid nitrogen. The harvested tissue is then stored at −80 C until RNA preparation.

The CMz036 (Lib3062) cDNA library is generated from maize husk tissue at the 8 week old plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from 8 week old plants. The husk is separated from the ear and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz037 (Lib3059) cDNA library is generated from maize pooled kernal at 12-15 days after pollienation plant development stage. Sample were collected from field grown material. Whole kernals from hand pollinated (control pollination) are harvested as whole ears and immediately frozen on dry ice. Kernels from 10-12 ears were pooled and ground together in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz039 (Lib3066) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz040 (Lib3067) cDNA library is generated from maize kernel tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold pollen. Five to eight days after controlled pollination. The ears are pulled and the kernels removed. The kernels are immediately frozen in liquid nitrogen. The harvested kernels tissue is then stored at −80° C. until RNA preparation. This sample represents gene expressed in early kernel development, during periods of cell division, amyloplast biogenesis and early carbon flow across the material to filial tissue.

The CMz041 (Lib3068) cDNA library is generated from maize pollen germinating silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants when the ear shoots are ready for fertilization at the silk emergence stage. The emerging silks are pollinated with an excess of pollen under controlled pollination conditions in the green house. Eighteen hours after pollination the silks are removed from the ears and immediately frozen in liquid nitrogen containers. This sample represents genes expressed in both pollen and silk tissue early in pollination. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz042 (Lib3069) cDNA library is generated from maize ear tissue excessively pollinated at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants and the ear shoots which are ready for fertilization are at the silk emergence stage. The immature ears are pollinated with an excess of pollen under controlled pollination conditions. Eighteen hours post-pollination, the ears are removed and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz044 (Lib3075) cDNA library is generated from maize microspore tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature anthers from 7 week old tassels. The immature anthers are first dissected from the 7 week old tassel with a scalpel on a glass slide covered with water. The microspores (immature pollen) are released into the water and are recovered by centrifugation. The microspore suspension is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz045 (Lib3076) cDNA library is generated from maize immature ear megaspore tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature ear (megaspore) obtained from 7 week old plants. The immature ears are harvested from the 7 week old plants and are approximately 2.5 to 3 cm in length. The kernels are removed from the cob immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz047 (Lib3078) cDNA library is generated from maize C0₂ treated high-exposure shoot tissue at the V10+ plant development stage. RX601 maize seeds are sterilized for i minute with a 10% clorox solution. The seeds are rolled in germination paper, and germinated in 0.5 mM calcium sulfate solution for two days ate 30° C. The seedlings are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium at a rate of 2-3 seedlings per pot. Twenty pots are placed into a high CO₂ environment (approximately 1000 ppm CO₂). Twenty plants were grown under ambient greenhouse CO₂ (approximately 450 ppm CO₂). Plants are watered daily before transplantation and three times a week after transplantation. Peters 20-20-20 fertilizer is also lightly applied. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. At ten days post planting, the shoots from both atmosphere are frozen in liquid nitrogen and lightly ground. The roots are washed in deionized water to remove the support media and the tissue is immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz048 (Lib3079) cDNA library is generated from maize basal endosperm transfer layer tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ maize plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence, to withhold the pollen. Kernels are harvested at 12 days post-pollination and placed on wet ice for dissection. The kernels are cross sectioned laterally, dissecting just above the pedicel region, including 1-2 mm of the lower endosperm and the basal endosperm transfer region. The pedicel and lower endosperm region containing the basal endosperm transfer layer is pooled and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz049(Lib3088) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately transferred to liquid nitrogen container. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz050 (Lib3114) cDNA library is generated from maize silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is beyond the 10-leaf development stage and the ear shoots are approximately 15-20 cm in length. The ears are pulled and silks are separated from the ears and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON001 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) total leaf tissue at the V4 plant development stage. Leaf tissue from 38, field grown V4 stage plants is harvested from the 4^(th) node. Leaf tissue is removed from the plants and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON002 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue at the V4 plant development stage. Root tissue from 76, field grown V4 stage plants is harvested. The root systems is cut from the soybean plant and washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON003 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON004 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledon tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON005 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after the start of imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post imbibition. At the 6 hours after imbibition stage, not all cotyledons have become fully hydrated and germination, or radicle protrusion, has not occurred. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON006 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledons tissue harvest 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nightime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post-imbibition. At the 6 hours after imbibition, not all cotyledons have become fully hydrated and germination or radicle protrusion, have not occurred. The seedlings are washed in water to remove soil, cotyledon harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON007 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days post-flowering. Seed pods from field grown plants are harvested 25 and 35 days after flowering and the seeds extracted from the pods. Approximately 4.4 g and 19.3 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON008 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested from 25 and 35 days post-flowering plants. Total leaf tissue is harvested from field grown plants. Approximately 19 g and 29 g of leaves are harvested from the fourth node of the plant 25 and 35 days post-flowering and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON009 cDNA library is generated from soybean cutlivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) pod and seed tissue harvested 15 days post-flowering. Pods from field grown plants are harvested 15 days post-flowering. Approximately 3 g of pod tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON010 cDNA library is generated from soybean cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) seed tissue harvested 40 days post-flowering. Pods from field grown plants are harvested 40 days post-flowering. Pods and seeds are separated, approximately 19 g of seed tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON011 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4^(th) node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON012 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue. Leaves from field grown plants are harvested from the fourth node 15 days post-flowering. Approximately 12 g of leaves are harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON013 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root and nodule tissue. Approximately, 28 g of root tissue from field grown plants is harvested 15 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON014 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days after flowering. Seed pods from field grown plants are harvested 15 days after flowering and the seeds extracted from the pods. Approximately 5 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON015 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 45 and 55 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 19 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON016 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately, 61 g and 38 g of root tissue from field grown plants is harvested 25 and 35 days post-flowering is harvested. The root system is cut from the soybean plant and washed with water to free it from the soil. The tissue is placed in 14ml polystyrene tubes and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON017 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately 28 g of root tissue from field grown plants is harvested 45 and 55 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON018 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 45 and 55 days post-flowering. Leaves from field grown plants are harvested 45 and 55 days after flowering from the fourth node. Approximately 27 g and 33 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON019 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON020 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 65 and 75 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 14 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON021 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in tissue culture at room temperature. At approximately 6 weeks post-germination, the plants are exposed to sterilized Soybean Cyst Nematode eggs. Infection is then allowed to progress for 10 days. After the 10 day infection process, the tissue is harvested. Agar from the culture medium and nematodes are removed and the root tissue is immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON022 (Lib3030) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) partially opened flower tissue. Partially to fully opened flower tissue is harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. A total of 3 g of flower tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON023 cDNA library is generated from soybean genotype BW211S Null (Tohoku University, Morioka, Japan) seed tissue harvested 15 and 40 days post-flowering. Seed pods from field grown plants are harvested 15 and 40 days post-flowering and the seeds extracted from the pods. Approximately 0.7 g and 14.2 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON024 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) internode-2 tissue harvested 18 days post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. The plants are grown in a greenhouse for 18 days after the start of imbibition at ambient temperature. Soil is checked and watered daily to maintain even moisture conditions. Stem tissue is harvested 18 days after the start of imbibition. The samples are divided into hypocotyl and internodes 1 through 5. The fifth internode contains some leaf bud material. Approximately 3 g of each sample is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON025 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 65 days post-flowering. Leaves are harvested from the fourth node of field grown plants 65 days post-flowering. Approximately 18.4 g of leaf tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

SOYMON026 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue harvested 65 and 75 days post-flowering. Approximately 27 g and 40 g of root tissue from field grown plants is harvested 65 and 75 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON027 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 days post-flowering. Seed pods from field grown plants are harvested 25 days post-flowering and the seeds extracted from the pods. Approximately 17 g of seeds are harvested from the seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON028 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed root tissue. The plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of development, water is withheld from half of the plant collection (drought stressed population). After 3 days, half of the plants from the drought stressed condition and half of the plants from the control population are harvested. After another 3 days (6 days post drought induction) the remaining plants are harvested. A total of 27 g and 40 g of root tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON029 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar PI07354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Late fall to early winter greenhouse grown plants are exposed to Soybean Cyst Nematode eggs. At 10 days post-infection, the plants are uprooted, rinsed briefly and the roots frozen in liquid nitrogen. Approximately 20 grams of root tissue is harvested from the infected plants. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON030 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) flower bud tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. A total of 100 mg of flower buds are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON031 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) carpel and stamen tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. Flowers are dissected to separate petals, sepals and reproductive structures (carpels and stamens). A total of 300 mg of carpel and stamen tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON032 cDNA library is prepared from the Asgrow cultivar A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry soybean seed meristem tissue. Surface sterilized seeds are germinated in liquid media for 24 hours. The seed axis is then excised from the barely germinating seed, placed on tissue culture media and incubated overnight at 20° C. in the dark. The supportive tissue is removed from the explant prior to harvest. Approximately 570 mg of tissue is harvested and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON033 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heat-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to an incubator set at 40° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance. Total RNA and poly A⁺ RNA is prepared from equal amounts of pooled tissue.

The SOYMON034 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) cold-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to a cold room set at 5° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance.

The SOYMON035 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed coat tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are harvested from mid to nearly full maturation (seed coats are not yellowing). The entire embryo proper is removed from the seed coat sample and the seed coat tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON036 cDNA library is generated from soybean cultivars PI171451, PI227687 and PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) insect challenged leaves. Plants from each of the three cultivars are grown in screenhouse conditions. The screenhouse is divided in half and one half of the screenhouse is infested with soybean looper and the other half infested with velvetbean caterpillar. A single leaf is taken from each of the representative plants at 3 different time points, 11 days after infestation, 2 weeks after infestation and 5 weeks after infestation and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Total RNA and poly A+ RNA is isolated from pooled tissue consisting of equal quantities of all 18 samples (3 genotypes×3 sample times×2 insect genotypes).

The SOYMON037 cDNA library is generated from soybean cultivar A3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) etiolated axis and radical tissue. Seeds are planted in moist vermiculite, wrapped and kept at room temperature in complete darkness until harvest. Etiolated axis and hypocotyl tissue is harvested at 2, 3 and 4 days post-planting. A total of 1 gram of each tissue type is harvested at 2, 3 and 4 days after planting and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON038 cDNA library is generated from soybean variety Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry seeds. Explants are prepared for transformation after germination of surface-sterilized seeds on solid tissue media. After 6 days, at 28° C. and 18 hours of light per day, the germinated seeds are cold shocked at 4° C. for 24 hours. Meristemic tissue and part of the hypocotyl is remove and cotyledon excised. The prepared explant is then wounded for Agrobacterium infection. The 2 grams of harvested tissue is frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The Soy51 (LIB3027) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy52 (LIB3028) cDNA library is generated from normalized flower DNA. Single stranded DNA representing approximately 1×10⁶ colony forming units of SOYMON022 harvested tissue is used as the starting material for normalization. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy53 (LIB3039) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling shoot apical meristem tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Apical tissue is harvested from seedling shoot meristem tissue, 7-8 days after the start of imbibition. The apex of each seedling is dissected to include the fifth node to the apical meristem. The fifth node corresponds to the third trifoliate leaf in the very early stages of development. Stipules completely envelop the leaf primordia at this time. A total of 200 mg of apical tissue is harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The Soy54 (LIB3040) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heart to torpedo stage embryo tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected and embryos removed from surrounding endosperm and maternal tissues. Embryos from globular to young torpedo stages (by corresponding analogy to Arabidopsis) are collected with a bias towards the middle of this spectrum. Embryos which are beginning to show asymmetric development of cotyledons are considered the upper developmental boundary for the collection and are excluded. A total of 12 mg embryo tissue is frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy55 (LIB3049) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) young seed tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected from very young pods (5 to 15 days after flowering). A total of 100 mg of seeds are harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy56 (LIB3029) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are not converted to double stranded form and represent a non-normalized seed pool for comparison to Soy51 cDNA libraries.

The Soy58 (LIB3050) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed root tissue subtracted from control root tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days root tissue from both drought stressed and control (watered regularly) plants are collected and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.).

The Soy59 (LIB3051) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) endosperm tissue. Seeds are germinated on paper towels under laboratory ambient light conditions. At 8, 10 and 14 hours after imbibition, the seed coats are harvested. The endosperm consists of a very thin layer of tissue affixed to the inside of the seed coat. The seed coat and endosperm are frozen immediately after harvest in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy60 (LIB3072) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed seed plus pod subtracted from control seed plus pod tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.).

The Soy61 (LIB3073) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is soaked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.). For this library's construction, the eighth fraction of the cDNA size fractionation step was used for ligation.

The Soy62 (LIB3074) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is soaked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.). For this library's construction, the ninth fraction of the cDNA size fractionation step was used for ligation.

The Soy65 (LIB3107) 07cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At the R3 stage of development, drought is imposed by withholding water. At 3, 4, 5 and 6 days, tissue is harvested and wilting is not obvious until the fourth day. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy66 (LIB3109) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) non-drought stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, control abscission layer tissue is harvested. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy67 (LIB3065) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy68 (LIB3052) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy69 (LIB3053) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) normalized leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4^(th) node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×10⁶ colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

Soy70 (LIB3055) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4^(th) node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy71 (LIB3056) cDNA library is generated from soybean cultivars Cristalina and FT108 (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy72 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.).

Soy73 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.).

The Soy76 (Lib3106) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid and arachidonic treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is soaked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.) in order to capture some of the smaller transcripts characteristic of antifungal proteins.

Soy77 (LIB3108) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1 % Tween-20. Plants are sprayed until runoff and the soil and the stem is soaked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 1 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad, Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector in order to capture some of the smaller transcripts characteristic of antifungal proteins.

The Lib9 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Leaf blades were cut with sharp scissors at seven weeks after planting. The tissue was immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib22 cDNA library is prepared from Arabidopsis thaliana Columbia ecotype, root tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems, floral buds appear, and a few flowers are starting to open. The 7-week old plants are rinsed intensively by tope water remove dirt from the roots, and blotted by paper towel. The tissues are immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Lib23 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, stem tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Stems were collected seven to eight weeks after planting by cutting the stems from the base and cutting the top of the plant to remove the floral tissue. The tissue was immediately frozen in liquid nitrogen and stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib24 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, flower bud tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flower buds are green and unopened and harvested about seven weeks after planting. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib25 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, open flower tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flowers are completely opened with all parts of floral structure observable, but no siliques are appearing. The tissue was immediately frozen in liquid nitrogen and stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib35 cDNA library of the present invention, was prepared from Arabidopsis thaliana Columbia ecotype leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems and floral buds appeared and a few flowers were starting to open. Leaf blades were collected by cutting with sharp scissors. The tissues were immediately frozen in liquid nitrogen and stored at −80° C. until use. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib146 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, immature seed tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. At approximately 7-8 weeks of age, the seeds are harvested. The seeds ranged in maturity from the smallest seeds that could be dissected from silques to just before starting to turn yellow in color. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)₂₅ (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library is normalized using a PCR-based protocol.

The Lib3032 (Lib80) cDNA libraries are generated from Brassica napus seeds harvested 30 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersgurg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis is carried out at 45° C.

The Lib3034 (Lib82) cDNA libraries are generated from Brassica napus seeds harvested 15 and 18 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib3099 cDNA library is generated by a subtraction procedure. The library contains cDNAs whose abundance is enriched in the Brassica napus 15 and 18 day after pollination seed tissues when compared to Brassica leaf tissues. The cDNA synthesis is performed on Brassica leaf RNA and Brassica RNA isolated from seeds harvested 15 and 18 days after pollination using a Smart PCR cDNA synthesis kit according to the manufacturers protocol (Clontech, Palo Alto, Calif. U.S.A.). The subtacted cDNA is generated using the Clontech PCR-Select subtaction kit according to the manufacturers protocol (Clontech, Palo Alto, Calif. U.S.A.). The subtacted cDNA was cloned into plasmid vector pCR2.1 according to the manufacturers protocol (Invitrogen, Carlsbad, Calif. U.S.A.).

The Lib3033 (Lib81) cDNA libraries are generated from from the Schizochytrium species cells. The Schizochytrium species cells are grown in liquid media until saturation. The culture is centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib47 cDNA library is generated from Euglena gracilus strain 753 (ATTC No. 30285, ATCC Manasas, Va. U.S.A.) grown in liquid culture. A liquid culture is innoculated with 1/10 volume of a previously-grown saturated culture, and the new culture for 4 days under near-anaerobic conditions (near-anaerobic cultures are not agitated, just gently swirled once a day) in the dark in 2× Beef (10 g/l bacto peptone, 4 g/l yeast extract, 2 g/l beef extract, 6 g/l glucose). The culture is then centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib44 cDNA library is generated from Phaeodactylum tricornatum grown in modified Jones medium for 3 days. The cells were harvested by centrifugation and the resulting pellet frozen immediately in liquid nitrogen. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Phaeodactylum cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45 degrees centigrade.

The LIB3036 genomic library is generated from Mycobacterium neoaurum US52 (ATCC No. 23072, ATCC, Manasas, Va. U.S.A.) cells. Mycobacterium neoaurum US52 is a gram-positive Actinomycete bacterium. Mycobacterium neoaurum US52 is genetically related to Mycobacterium tuberculosis, but there is no reason to believe that it is a primary pathogen. It normally is saprophytic, i.e. it lives in soil and gets nutrients from decaying matter. Genomic DNA obtained from Mycobacterium neoaurum US52 is digested for various times with the restriction enzyme Sau3A. The DNA fractions are size-separated on an agarose gel, and the first fraction wherein most of the partially-digested fragments are about 10 kB is used to isolated fragments in the range of 2-3 kB. For LIB3036, the 2-3 kB DNA is cloned into vector pRY401 (Invitrogen, Carlsbad, Calif. U.S.A.). The vector pZERO-2 (Invitrogen, Carlsbad, Calif. U.S.A.). is used for the construction of LIB3104.

The stored RNA is purified using Trizol reagent from Life Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially as recommended by the manufacturer. Poly A+ RNA (mRNA) is purified using magnetic oligo dT beads essentially as recommended by the manufacturer (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).

Construction of plant cDNA libraries is well-known in the art and a number of cloning strategies exist. A number of cDNA library construction kits are commercially available. The Superscript™ Plasmid System for cDNA synthesis and Plasmid Cloning (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.) is used, following the conditions suggested by the manufacturer.

Normalized libraries are made using essentially the Soares procedure (Soares et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:9228-9232 (1994), the entirety of which is herein incorporated by reference). This approach is designed to reduce the initial 10,000-fold variation in individual cDNA frequencies to achieve abundances within one order of magnitude while maintaining the overall sequence complexity of the library. In the normalization process, the prevalence of high-abundance cDNA clones decreases dramatically, clones with mid-level abundance are relatively unaffected and clones for rare transcripts are effectively increased in abundance.

EXAMPLE 2

The cDNA libraries are plated on LB agar containing the appropriate antibiotics for selection and incubated at 37° for a sufficient time to allow the growth of individual colonies. Single colonies are individually placed in each well of a 96-well microtiter plates containing LB liquid including the selective antibiotics. The plates are incubated overnight at approximately 37° C. with gentle shaking to promote growth of the cultures. The plasmid DNA is isolated from each clone using Qiaprep plasmid isolation kits, using the conditions recommended by the manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).

Template plasmid DNA clones are used for subsequent sequencing. For sequencing, the ABI PRISM dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq® DNA Polymerase, FS, is used (PE Applied Biosystems, Foster City, Calif. U.S.A.).

EXAMPLE 3

Nucleic acid sequences that encode for the following proteins: triose phosphate isomerase, fructose 1,6-bisphosphate aldolase, fructose 1,6-bisphosphate, fructose 6-phosphate 2-kinase, phosphoglucoisomerase, vacuolar H⁺ translocating-pyrophosphatase, pyrophosphate-dependent fructose-6-phosphate phosphotransferase, invertase, sucrose synthase, hexokinase, fructokinase, NDP-kinase, glucose-6-phosphate 1-dehydrogenase, phosphoglucomutase and UDP-glucose pyrophophorylase are identified from the Monsanto EST PhytoSeq database using TBLASTN (default values)(TBLASTN compares a protein query against the six reading frames of a nucleic acid sequence). Matches found with BLAST P values equal or less than 0.001 (probability) or BLAST Score of equal or greater than 90 are classified as hits. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

In addition, the GenBank database is searched with BLASTN and BLASTX (default values) using ESTs as queries. EST that pass the hit probability threshold of 10e⁻⁸ for the following enzymes are combined with the hits generated by using TBLASTN (described above) and classified by enzyme (see Table A below).

A cluster refers to a set of overlapping clones in the PhytoSeq database. Such an overlapping relationship among clones is designated as a “cluster” when BLAST scores from pairwise sequence comparisons of the member clones meets a predetermined minimum value or product score of 50 or more (Product Score=(BLAST SCORE×Percentage Identity)/(5×minimum [length (Seq1), length (Seq2)]))

Since clusters are formed on the basis of single-linkage relationships, it is possible for two non-overlapping clones to be members of the same cluster if, for instance, they both overlap a third clone with at least the predetermined minimum BLAST score (stringency). A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. Clones grouped in a cluster in most cases represent a contiguous sequence.

TABLE A* Seq No. Cluster ID CloneID Library NCBI gi Method Score P-value % Ident MAIZE TRIOSE PHOSPHATE ISOMERASE 1 −700019675 700019675H1 SATMON001 g546735 BLASTX 134 1e−11 78 2 −700073894 700073894H1 SATMON007 g609261 BLASTN 257 1e−10 84 3 −700167260 700167260H1 SATMON013 g609261 BLASTN 644 1e−44 79 4 −700380595 700380595H1 SATMON021 g609261 BLASTN 1121 1e−84 87 5 −700449667 700449667H1 SATMON028 g217973 BLASTN 204 1e−18 93 6 −700449720 700449720H2 SATMON028 g217973 BLASTN 216 1e−18 88 7 −700570661 700570661H1 SATMON030 g168647 BLASTX 131 1e−11 88 8 −700616770 700616770H1 SATMON033 g407525 BLASTX 149 1e−13 83 9 −701170944 701170944H1 SATMONN05 g217921 BLASTX 188 1e−20 53 10 11337 700337974H1 SATMON020 g256119 BLASTN 535 1e−61 78 11 11337 700027829H1 SATMON003 g256119 BLASTN 726 1e−51 80 12 126 700050046H1 SATMON003 g1785947 BLASTN 440 1e−26 92 13 282 700077320H1 SATMON007 g217973 BLASTN 666 1e−108 97 14 282 700104541H1 SATMON010 g217973 BLASTN 631 1e−106 97 15 282 700047476H1 SATMON003 g217973 BLASTN 648 1e−105 97 16 282 700211559H1 SATMON016 g217973 BLASTN 525 1e−104 97 17 282 700073553H1 SATMON007 g217973 BLASTN 981 1e−103 98 18 282 700613011H1 SATMON033 g217973 BLASTN 552 1e−102 98 19 282 700352119H1 SATMON023 g217973 BLASTN 666 1e−101 97 20 282 700088148H1 SATMON011 g217973 BLASTN 666 1e−100 98 21 282 700351626H1 SATMON023 g217973 BLASTN 401 1e−99 98 22 282 700240096H1 SATMON010 g217973 BLASTN 666 1e−98 97 23 282 700083660H1 SATMON011 g217973 BLASTN 666 1e−97 99 24 282 700208721H1 SATMON016 g217973 BLASTN 497 1e−96 98 25 282 700203144H1 SATMON003 g217973 BLASTN 511 1e−96 96 26 282 700430425H1 SATMONN01 g217973 BLASTN 666 1e−96 98 27 282 700206091H1 SATMON003 g217973 BLASTN 497 1e−94 97 28 282 700077017H1 SATMON007 g217973 BLASTN 614 1e−93 93 29 282 700618792H1 SATMON034 g217973 BLASTN 546 1e−92 96 30 282 700572532H1 SATMON030 g407524 BLASTN 1212 1e−92 84 31 282 700106512H1 SATMON010 g217973 BLASTN 632 1e−91 97 32 282 700195031H1 SATMON014 g217973 BLASTN 471 1e−90 97 33 282 700168131H1 SATMON013 g217973 BLASTN 497 1e−89 98 34 282 700197039H1 SATMON014 g217973 BLASTN 546 1e−89 98 35 282 700572688H1 SATMON030 g169820 BLASTN 1114 1e−89 85 36 282 700021313H1 SATMON001 g217973 BLASTN 913 1e−87 97 37 282 700452417H1 SATMON028 g217973 BLASTN 425 1e−86 95 38 282 700346119H1 SATMON021 g217973 BLASTN 444 1e−86 96 39 282 700082359H1 SATMON011 g217973 BLASTN 542 1e−86 93 40 282 700240042H1 SATMON010 g217973 BLASTN 596 1e−86 97 41 282 700030064H1 SATMON003 g217973 BLASTN 587 1e−85 94 42 282 700615185H1 SATMON033 g217973 BLASTN 430 1e−84 98 43 282 700196125H1 SATMON014 g217973 BLASTN 581 1e−84 100 44 282 700243429H1 SATMON010 g217973 BLASTN 632 1e−84 97 45 282 700474112H1 SATMON025 g217973 BLASTN 570 1e−83 98 46 282 700572282H1 SATMON030 g407524 BLASTN 838 1e−83 82 47 282 700622238H1 SATMON034 g169820 BLASTN 917 1e−80 86 48 282 700095609H1 SATMON008 g169820 BLASTN 1067 1e−80 82 49 282 700218886H1 SATMON011 g217973 BLASTN 551 1e−79 93 50 282 700018688H1 SATMON001 g217973 BLASTN 1066 1e−79 99 51 282 700049775H1 SATMON003 g217973 BLASTN 362 1e−78 91 52 282 700575972H1 SATMON030 g169820 BLASTN 894 1e−78 79 53 282 700215519H1 SATMON016 g217973 BLASTN 497 1e−76 97 54 282 700161120H1 SATMON012 g217973 BLASTN 622 1e−76 98 55 282 700581760H1 SATMON031 g217973 BLASTN 533 1e−75 90 56 282 700104672H1 SATMON010 g169820 BLASTN 1012 1e−75 83 57 282 700346053H1 SATMON021 g169820 BLASTN 1012 1e−75 83 58 282 701166592H1 SATMONN04 g217973 BLASTN 661 1e−74 95 59 282 700968667H1 SATMONN04 g217973 BLASTN 497 1e−73 92 60 282 700205627H1 SATMON003 g217973 BLASTN 666 1e−73 99 61 282 700029005H1 SATMON003 g169820 BLASTN 979 1e−72 85 62 282 700476479H1 SATMON025 g169820 BLASTN 554 1e−71 84 63 282 700050148H1 SATMON003 g169820 BLASTN 608 1e−70 83 64 282 700259846H1 SATMON017 g217973 BLASTN 283 1e−69 94 65 282 700344093H1 SATMON021 g169820 BLASTN 934 1e−69 83 66 282 700082327H1 SATMON011 g169820 BLASTN 943 1e−69 85 67 282 700020156H1 SATMON001 g217973 BLASTN 420 1e−68 99 68 282 700577714H1 SATMON031 g169820 BLASTN 928 1e−68 85 69 282 700104904H1 SATMON010 g169820 BLASTN 913 1e−67 84 70 282 700104685H1 SATMON010 g169820 BLASTN 897 1e−66 84 71 282 700053463H1 SATMON009 g169820 BLASTN 907 1e−66 85 72 282 700171639H1 SATMON013 g217973 BLASTN 401 1e−65 98 73 282 700574233H1 SATMON030 g169820 BLASTN 651 1e−65 83 74 282 700262653H1 SATMON017 g169820 BLASTN 877 1e−64 84 75 282 700456738H1 SATMON029 g169820 BLASTN 877 1e−64 84 76 282 700611806H1 SATMON022 g169820 BLASTN 877 1e−64 83 77 282 700381177H1 SATMON023 g169820 BLASTN 884 1e−64 84 78 282 700103347H1 SATMON010 g169820 BLASTN 861 1e−63 84 79 282 700103605H1 SATMON010 g169820 BLASTN 868 1e−63 84 80 282 700578536H1 SATMON031 g169820 BLASTN 856 1e−62 84 81 282 700258606H1 SATMON017 g169820 BLASTN 807 1e−61 83 82 282 700335703H1 SATMON019 g217973 BLASTN 376 1e−60 90 83 282 700351044H1 SATMON023 g169820 BLASTN 471 1e−59 83 84 282 700346364H1 SATMON021 g169820 BLASTN 813 1e−59 85 85 282 700619037H1 SATMON034 g169820 BLASTN 814 1e−59 84 86 282 700465160H1 SATMON025 g169820 BLASTN 751 1e−57 84 87 282 700235687H1 SATMON010 g169820 BLASTN 791 1e−57 82 88 282 700105645H1 SATMON010 g169820 BLASTN 793 1e−57 83 89 282 700082237H1 SATMON011 g169820 BLASTN 793 1e−57 84 90 282 700261906H1 SATMON017 g169820 BLASTN 796 1e−57 83 91 282 700456154H1 SATMON029 g169820 BLASTN 799 1e−57 84 92 282 700047696H1 SATMON003 g169820 BLASTN 561 1e−56 83 93 282 700449905H1 SATMON028 g169820 BLASTN 788 1e−56 84 94 282 700336106H1 SATMON019 g217973 BLASTN 325 1e−55 92 95 282 700381867H1 SATMON023 g2529386 BLASTN 422 1e−55 97 96 282 700051335H1 SATMON003 g169820 BLASTN 608 1e−55 83 97 282 700050988H1 SATMON003 g169820 BLASTN 768 1e−55 86 98 282 700029471H1 SATMON003 g169820 BLASTN 772 1e−55 84 99 282 700106806H1 SATMON010 g169820 BLASTN 773 1e−55 84 100 282 700071749H1 SATMON007 g217973 BLASTN 362 1e−54 85 101 282 700207607H1 SATMON016 g217973 BLASTN 362 1e−54 85 102 282 700573465H2 SATMON030 g169820 BLASTN 753 1e−54 86 103 282 700220908H1 SATMON011 g169820 BLASTN 758 1e−54 84 104 282 700467719H1 SATMON025 g169820 BLASTN 761 1e−54 85 105 282 700456018H1 SATMON029 g169820 BLASTN 764 1e−54 81 106 282 700453767H1 SATMON029 g217973 BLASTN 296 1e−52 94 107 282 700026118H1 SATMON003 g217973 BLASTN 341 1e−52 93 108 282 700026760H1 SATMON003 g217973 BLASTN 421 1e−52 99 109 282 700029525H1 SATMON003 g169820 BLASTN 738 1e−52 85 110 282 700457972H1 SATMON029 g169820 BLASTN 723 1e−51 85 111 282 700455866H1 SATMON029 g169820 BLASTN 726 1e−51 84 112 282 700165290H1 SATMON013 g169820 BLASTN 726 1e−51 84 113 282 700351190H1 SATMON023 g169820 BLASTN 672 1e−50 81 114 282 700154095H1 SATMON007 g169820 BLASTN 696 1e−49 84 115 282 700450438H1 SATMON028 g217973 BLASTN 430 1e−48 99 116 282 700044892H1 SATMON004 g169820 BLASTN 683 1e−48 85 117 282 700185095H1 SATMON014 g169820 BLASTN 673 1e−47 84 118 282 700575506H1 SATMON030 g169820 BLASTN 680 1e−47 83 119 282 700161966H1 SATMON012 g217973 BLASTN 335 1e−46 98 120 282 700343401H1 SATMON021 g169820 BLASTN 426 1e−45 77 121 282 700152354H1 SATMON007 g169820 BLASTN 653 1e−45 84 122 282 701164924H1 SATMONN04 g169820 BLASTN 397 1e−44 84 123 282 700346896H1 SATMON021 g169820 BLASTN 496 1e−42 84 124 282 700210157H1 SATMON016 g169820 BLASTN 617 1e−42 84 125 282 700383103H1 SATMON024 g169820 BLASTN 531 1e−41 84 126 282 701158829H1 SATMONN04 g407524 BLASTN 549 1e−40 80 127 282 700619883H1 SATMON034 g217973 BLASTN 325 1e−38 99 128 282 700168219H1 SATMON013 g169820 BLASTN 540 1e−36 83 129 282 700155210H1 SATMON007 g169820 BLASTN 545 1e−36 83 130 282 700334861H1 SATMON019 g169820 BLASTN 484 1e−31 82 131 282 700355663H1 SATMON024 g217973 BLASTN 213 1e−30 88 132 282 700074764H1 SATMON007 g546734 BLASTN 387 1e−27 84 133 282 700621934H1 SATMON034 g217973 BLASTN 430 1e−26 100 134 282 700802084H1 SATMON036 g217973 BLASTN 270 1e−24 98 135 3039 700620444H1 SATMON034 g1785947 BLASTN 473 1e−56 75 136 3039 700356205H1 SATMON024 g1785947 BLASTN 332 1e−32 72 137 3039 700215549H1 SATMON016 g414549 BLASTN 443 1e−26 72 138 3039 700620318H1 SATMON034 g556171 BLASTX 214 1e−25 79 139 3039 700028742H1 SATMON003 g556171 BLASTX 156 1e−20 86 140 3039 700150060H1 SATMON007 g556171 BLASTX 181 1e−17 89 141 3039 700448477H1 SATMON027 g556171 BLASTX 137 1e−12 85 142 3039 700336489H1 SATMON019 g556171 BLASTX 126 1e−10 81 143 3414 700099709H1 SATMON009 g609261 BLASTN 600 1e−49 84 144 3414 700075837H1 SATMON007 g609261 BLASTN 494 1e−41 84 145 3414 700045678H1 SATMON004 g609261 BLASTN 340 1e−29 73 146 3414 700097852H1 SATMON009 g609261 BLASTN 436 1e−27 84 147 3414 700053342H1 SATMON009 g609261 BLASTN 346 1e−25 73 148 3414 700041954H1 SATMON004 g609261 BLASTN 340 1e−24 82 149 3414 700217471H1 SATMON016 g609261 BLASTN 265 1e−21 71 150 3414 700264437H1 SATMON017 g609261 BLASTN 231 1e−17 69 151 3414 700218371H1 SATMON016 g609261 BLASTN 156 1e−10 68 152 5593 700381686H1 SATMON023 g609261 BLASTN 534 1e−44 89 153 5593 700356082H1 SATMON024 g609261 BLASTN 246 1e−24 90 154 5593 700622077H1 SATMON034 g609261 BLASTN 292 1e−20 86 155 5593 700470822H1 SATMON025 g609262 BLASTX 134 1e−11 79 156 6525 700083139H1 SATMON011 g256119 BLASTN 880 1e−64 76 157 6525 700205474H1 SATMON003 g169820 BLASTN 849 1e−62 77 158 6991 700336856H1 SATMON019 g609261 BLASTN 1131 1e−85 85 159 6991 700042717H1 SATMON004 g609261 BLASTN 1028 1e−76 85 160 6991 700379491H1 SATMON020 g609261 BLASTN 995 1e−74 81 161 6991 700156635H1 SATMON012 g609261 BLASTN 877 1e−64 84 162 6991 700046340H1 SATMON004 g609261 BLASTN 852 1e−62 84 163 6991 700081869H1 SATMON011 g609261 BLASTN 266 1e−14 80 164 6991 700426102H1 SATMONN01 g806312 BLASTX 134 1e−13 89 165 7384 700613626H1 SATMON033 g609261 BLASTN 920 1e−87 85 166 7384 700101506H1 SATMON009 g609261 BLASTN 1124 1e−84 85 167 7384 700206445H1 SATMON003 g609261 BLASTN 987 1e−73 79 168 7384 700220160H1 SATMON011 g609261 BLASTN 878 1e−64 85 169 −L1431527 LIB143-004- LIB143 g217973 BLASTN 290 1e−13 93 Q1-E1-C5 170 −L30613868 LIB3061-017- LIB3061 g217973 BLASTN 182 1e−13 70 Q1-K1-C9 171 −L30623620 LIB3062-034- LIB3062 g609261 BLASTN 599 1e−39 74 Q1-K1-A8 172 −L361705 LIB36-021- LIB36 g609261 BLASTN 266 1e−14 80 Q1-E1-E7 173 23992 LIB3062-056- LIB3062 g1200507 BLASTX 285 1e−64 61 Q1-K1-F9 174 282 LIB3067-047- LIB3067 g217973 BLASTN 1076 1e−164 96 Q1-K1-H2 175 282 LIB3067-055- LIB3067 g217973 BLASTN 1076 1e−133 93 Q1-K1-G8 176 282 LIB3067-059- LIB3067 g169820 BLASTN 1401 1e−115 84 Q1-K1-D10 177 282 LIB3067-027- LIB3067 g407524 BLASTN 995 1e−113 83 Q1-K1-B10 178 282 LIB189-032- LIB189 g217973 BLASTN 629 1e−111 93 Q1-E1-H2 179 282 LIB3059-023- LIB3059 g407524 BLASTN 1436 1e−111 83 Q1-K1-A7 180 282 LIB3069-016- LIB3069 g169820 BLASTN 1301 1e−107 81 Q1-K1-D9 181 282 LIB143-006- LIB143 g169820 BLASTN 1373 1e−105 84 Q1-E1-A8 182 282 LIB3068-054- LIB3068 g169820 BLASTN 1327 1e−102 82 Q1-K1-C11 183 282 LIB3067-034- LIB3067 g407524 BLASTN 1321 1e−101 83 Q1-K1-B7 184 282 LIB143-031- LIB143 g169820 BLASTN 1311 1e−100 84 Q1-E1-E5 185 282 LIB3069-055- LIB3069 g169820 BLASTN 1046 1e−97 75 Q1-K1-H12 186 282 LIB3061-027- LIB3061 g169820 BLASTN 936 1e−96 83 Q1-K1-A8 187 282 LIB3078-008- LIB3078 g169820 BLASTN 1210 1e−92 82 Q1-K1-E5 188 282 LIB3066-027- LIB3066 g407524 BLASTN 1196 1e−91 82 Q1-K1-E1 189 282 LIB3067-032- LIB3067 g169820 BLASTN 1122 1e−84 84 Q1-K1-E5 190 282 LIB3078-029- LIB3078 g169820 BLASTN 827 1e−83 82 Q1-K1-F7 191 282 LIB3061-006- LIB3061 g169820 BLASTN 1091 1e−82 78 Q1-K1-B7 192 282 LIB143-048- LIB143 g169820 BLASTN 644 1e−74 75 Q1-E1-F8 193 282 LIB3078-033- LIB3078 g169820 BLASTN 584 1e−73 79 Q1-K1-B10 194 282 LIB3069-046- LIB3069 g169820 BLASTN 819 1e−59 79 Q1-K1-C4 195 282 LIB3061-049- LIB3061 g169820 BLASTN 587 1e−47 80 Q1-K1-H2 196 282 LIB143-029- LIB143 g169820 BLASTN 679 1e−47 84 Q1-E1-G4 197 282 LIB84-027- LIB84 g169820 BLASTN 613 1e−46 78 Q1-E1-E5 198 282 LIB3062-00l- LIB3062 g169820 BLASTN 507 1e−33 80 Q1-K2-F7 199 282 LIB3066-014- LIB3066 g169820 BLASTN 385 1e−25 76 Q1-K1-H11 200 29645 LIB3069-014- LIB3069 g168647 BLASTX 131 1e−27 34 Q1-K1-C11 201 29645 LIB3069-013- LIB3069 g168647 BLASTX 124 1e−24 33 Q1-K1-C11 202 3039 LIB3062-045- LIB3062 g1785947 BLASTN 1119 1e−84 72 Q1-K1-F6 203 5593 LIB3067-045- LIB3067 g609261 BLASTN 702 1e−58 75 Q1-K1-E5 204 6991 LIB3059-026- LIB3059 g609261 BLASTN 1493 1e−115 84 Q1-K1-G9 205 6991 LIB3078-049- LIB3078 g609261 BLASTN 747 1e−55 83 Q1-K1-E4 206 7384 LIB3062-034- LIB3062 g609261 BLASTN 1351 1e−107 85 Q1-K1-A4 MAIZE FRUCTOSE 1,6-BISPHOSPHATE ALDOLASE 207 −700026544 700026544H1 SATMON003 g22144 BLASTN 215 1e−30 88 208 −700073329 700073329H1 SATMON007 g22144 BLASTN 590 1e−89 95 209 −700151987 700151987H1 SATMON007 g22144 BLASTN 212 1e−8 78 210 −700206575 700206575H1 SATMON003 g22144 BLASTN 1009 1e−109 96 211 −700333727 700333727H1 SATMON019 g1217893 BLASTX 154 1e−16 61 212 −700429795 700429795H1 SATMONN01 g1619605 BLASTX 102 1e−16 77 213 −700804137 700804137H1 SATMON036 g22144 BLASTN 742 1e−52 92 214 1182 700449930H1 SATMON028 g22632 BLASTN 856 1e−62 79 215 1182 701185559H1 SATMONN06 g22632 BLASTN 793 1e−57 79 216 1182 700203130H1 SATMON003 g22632 BLASTN 799 1e−57 78 217 1182 700083459H1 SATMON011 g22632 BLASTN 800 1e−57 76 218 1182 700465449H1 SATMON025 g22632 BLASTN 405 1e−50 76 219 1182 701165344H1 SATMONN04 g22632 BLASTN 326 1e−29 78 220 1182 700427538H1 SATMONN01 g438275 BLASTX 96 1e−9 88 221 38 700224356H1 SATMON011 g22144 BLASTN 1290 1e−98 96 222 38 700048169H1 SATMON003 g22144 BLASTN 528 1e−72 98 223 38 700616610H1 SATMON033 g22144 BLASTN 278 1e−31 91 224 38 700355765H1 SATMON024 g20204 BLASTX 141 1e−12 96 225 6547 700194431H1 SATMON014 g2636513 BLASTX 181 1e−17 47 226 6547 700469777H1 SATMON025 g2636513 BLASTX 174 1e−16 48 227 8494 700425929H1 SATMONN01 g927507 BLASTX 67 1e−11 89 228 −L30603643 LIB3060-046- LIB3060 g169037 BLASTX 155 1e−44 66 Q1-K1-G7 229 1182 LIB3079-006- LIB3079 g22632 BLASTN 598 1e−39 65 Q1-K1-H8 230 28633 LIB3062-015- LIB3062 g1208898 BLASTX 116 1e−24 45 Q1-K1-G12 231 38 LIB3061-025- LIB3061 g22144 BLASTN 895 1e−133 94 Q1-K1-C9 232 38 LIB3059-020- LIB3059 g22144 BLASTN 745 1e−53 98 Q1-K1-H3 MAIZE FRUCTOSE-1,6-BISPHOSPHATASE 233 −700262935 700262935H1 SATMON017 g3041775 BLASTX 184 1e−18 94 234 −700432173 700432173H1 SATMONN01 g1790679 BLASTX 123 1e−16 56 235 −700455709 700455709H1 SATMON029 g3041776 BLASTN 597 1e−40 85 236 −700573083 700573083H1 SATMON030 g3041775 BLASTX 69 1e−10 64 237 12846 700101851H1 SATMON009 g3041776 BLASTN 1312 1e−100 91 238 12846 700101541H1 SATMON009 g3041776 BLASTN 1252 1e−95 90 239 12846 700581510H1 SATMON031 g3041776 BLASTN 872 1e−82 90 240 15627 700046054H1 SATMON004 g21736 BLASTN 1213 1e−92 91 241 15627 700421605H1 SATMONN01 g3041776 BLASTN 664 1e−77 90 242 15627 700445495H1 SATMON027 g21736 BLASTN 1004 1e−74 84 243 15627 700042188H1 SATMON004 g3041776 BLASTN 875 1e−64 88 244 16870 700100752H1 SATMON009 g3041776 BLASTN 257 1e−33 75 245 16870 700044805H1 SATMON004 g3041776 BLASTN 194 1e−14 76 246 16870 700099217H1 SATMON009 g21736 BLASTN 246 1e−9 59 247 5480 700442189H1 SATMON026 g3041774 BLASTN 536 1e−54 93 248 8243 700264654H1 SATMON017 g3041774 BLASTN 942 1e−69 84 249 8243 700479624H1 SATMON034 g3041774 BLASTN 902 1e−66 82 250 8243 700448974H1 SATMON028 g3041774 BLASTN 876 1e−64 84 251 −L1485381 LIB148-057- LIB148 g440591 BLASTX 80 1e−30 63 Q1-E1-E6 252 −L30662839 LIB3066-035- LIB3066 g3041774 BLASTN 215 1e−15 77 Q1-K1-F11 253 −L362913 LIB36-013- LIB36 g3041776 BLASTN 937 1e−69 88 Q1-E1-D10 254 −L832444 LIB83-005- LIB83 g3041776 BLASTN 575 1e−37 93 Q1-E1-D2 255 12846 LIB83-008- LIB83 g3041776 BLASTN 1610 1e−135 92 Q1-E1-A8 256 12846 LIB3078-003- LIB3078 g3041776 BLASTN 873 1e−98 93 Q1-K1-C7 257 16870 LIB3060-052- LIB3060 g21736 BLASTN 377 1e−66 70 Q1-K1-D11 258 26002 LIB83-008- LIB83 g3041776 BLASTN 378 1e−20 86 Q1-E1-B10 MAIZE FRUCTOSE-6-PHOSPHATE,2-KINASE 259 −700093724 700093724H1 SATMON008 g3170230 BLASTX 123 1e−21 53 260 −700099547 700099547H1 SATMON009 g3309582 BLASTN 630 1e−43 80 261 −700100682 700100682H1 SATMON009 g3170230 BLASTX 269 1e−39 65 262 −700173085 700173085H1 SATMON013 g2286154 BLASTN 1165 1e−88 100 263 −700217623 700217623H1 SATMON016 g3170229 BLASTN 593 1e−40 73 264 −700219340 700219340H1 SATMON011 g3170230 BLASTX 190 1e−20 56 265 −700265353 700265353H1 SATMON017 g2286154 BLASTN 1268 1e−107 98 266 −700379777 700379777H1 SATMON021 g3309582 BLASTN 905 1e−66 76 267 −700620963 700620963H1 SATMON034 g2286154 BLASTN 376 1e−52 85 268 −701159590 701159590H1 SATMONN04 g3309582 BLASTN 682 1e−48 73 269 20094 700209789H1 SATMON016 g2286154 BLASTN 1093 1e−96 92 270 20094 700550375H1 SATMON022 g3309582 BLASTN 780 1e−58 81 271 29193 700021150H1 SATMON001 g2286154 BLASTN 466 1e−75 92 272 −L30593297 LIB3059-029- LIB3059 g2286154 BLASTN 401 1e−22 70 Q1-K1-B3 273 −L30614892 LIB3061-021- LIB3061 g2286154 BLASTN 469 1e−38 79 Q1-K1-G9 274 −L30623700 LIB3062-031- LIB3062 g3170229 BLASTN 230 1e−10 70 Q1-K1-E8 275 29193 LIB83-007- LIB83 g2286154 BLASTN 595 1e−113 90 Q1-E1-C11 MAIZE PHOSPHOGLUCOISOMERASE 276 −700086021 700086021H1 SATMON011 g1100771 BLASTX 225 1e−28 51 277 −700169489 700169489H1 SATMON013 g1100771 BLASTX 152 1e−13 59 278 −700222638 700222638H1 SATMON011 g1100771 BLASTX 256 1e−28 60 279 −700445574 700445574H1 SATMON027 g1100771 BLASTX 143 1e−12 54 280 −700475232 700475232H1 SATMON025 g596022 BLASTN 845 1e−61 90 281 −700612774 700612774H1 SATMON033 g596022 BLASTN 1574 1e−122 95 282 14393 700222547H1 SATMON011 g1100771 BLASTX 239 1e−25 60 283 14393 700220357H1 SATMON011 g1100771 BLASTX 218 1e−23 68 284 14393 700050317H1 SATMON003 g1100771 BLASTX 120 1e−22 63 285 14393 700163544H1 SATMON013 g1100771 BLASTX 214 1e−22 62 286 15724 700207164H1 SATMON017 g1100771 BLASTX 135 1e−17 67 287 15724 700552402H1 SATMON022 g1100771 BLASTX 135 1e−11 60 288 15724 700086085H1 SATMON011 g1100771 BLASTX 137 1e−11 45 289 20643 700577051H1 SATMON031 g1100771 BLASTX 241 1e−26 66 290 20643 700201592H1 SATMON003 g1100771 BLASTX 113 1e−19 45 291 20643 700576644H1 SATMON030 g1100771 BLASTX 113 1e−17 43 292 2351 700208928H1 SATMON016 g1100771 BLASTX 274 1e−43 73 293 2351 700240758H1 SATMON010 g1100771 BLASTX 283 1e−43 79 294 2351 700352502H1 SATMON023 g1100771 BLASTX 197 1e−36 70 295 2351 700581930H1 SATMON031 g1100771 BLASTX 164 1e−34 72 296 2351 700028642H1 SATMON003 g1100771 BLASTX 294 1e−33 65 297 2351 700106092H1 SATMON010 g1100771 BLASTX 294 1e−33 62 298 2351 700082102H1 SATMON011 g1100771 BLASTX 300 1e−33 62 299 2351 700083446H1 SATMON011 g1100771 BLASTX 274 1e−30 65 300 2351 700580585H1 SATMON031 g1100771 BLASTX 163 1e−29 69 301 2351 700550608H1 SATMON022 g1100771 BLASTX 265 1e−29 61 302 2351 700106079H1 SATMON010 g1100771 BLASTX 261 1e−28 54 303 2351 700244248H1 SATMON010 g1100771 BLASTX 238 1e−25 67 304 2351 700152233H1 SATMON007 g1100771 BLASTX 167 1e−22 72 305 2351 700455043H1 SATMON029 g1100771 BLASTX 168 1e−21 68 306 2351 700615809H1 SATMON033 g1100771 BLASTX 207 1e−21 66 307 2351 701165320H1 SATMONN04 g1100771 BLASTX 122 1e−14 63 308 32930 700042996H1 SATMON004 g596022 BLASTN 476 1e−95 98 309 4222 700222539H1 SATMON011 g596022 BLASTN 1160 1e−87 100 310 4222 700104023H1 SATMON010 g596022 BLASTN 1060 1e−84 100 311 4222 700101580H1 SATMON009 g596022 BLASTN 871 1e−74 99 312 4222 700473395H1 SATMON025 g596022 BLASTN 368 1e−46 95 313 4222 700800179H1 SATMON036 g596022 BLASTN 240 1e−11 100 314 8858 700221523H1 SATMON011 g1100771 BLASTX 278 1e−31 59 315 895 700100965H1 SATMON009 g596022 BLASTN 1611 1e−125 99 316 895 700620985H1 SATMON034 g596022 BLASTN 1418 1e−114 98 317 895 700082062H1 SATMON011 g596022 BLASTN 1365 1e−110 97 318 895 700573782H1 SATMON030 g596022 BLASTN 920 1e−107 98 319 895 700236138H1 SATMON010 g596022 BLASTN 1395 1e−107 100 320 895 700086336H1 SATMON011 g596022 BLASTN 1370 1e−105 100 321 895 700801467H1 SATMON036 g596022 BLASTN 1249 1e−99 95 322 895 700801458H1 SATMON036 g596022 BLASTN 1245 1e−98 100 323 895 700475024H1 SATMON025 g596022 BLASTN 1162 1e−97 93 324 895 700243164H1 SATMON010 g596022 BLASTN 1105 1e−96 100 325 895 700804665H1 SATMON036 g596022 BLASTN 1266 1e−96 99 326 895 700021931H1 SATMON001 g596022 BLASTN 1126 1e−84 99 327 895 700805540H1 SATMON036 g596022 BLASTN 776 1e−55 99 328 895 700172576H1 SATMON013 g596022 BLASTN 571 1e−38 98 329 895 700105116H1 SATMON010 g596022 BLASTN 558 1e−37 99 330 895 700472931H1 SATMON025 g596022 BLASTN 379 1e−31 97 331 20643 LIB3069-009- LIB3069 g1100771 BLASTX 215 1e−44 50 Q1-K1-B3 332 2351 LIB3079-007- LIB3079 g1100771 BLASTX 304 1e−77 72 Q1-K1-C11 333 32930 LIB189-001- LIB189 g596022 BLASTN 794 1e−115 95 Q1-E1-E4 334 4222 LIB3079-001- LIB3079 g596022 BLASTN 1132 1e−101 89 Q1-K1-H7 335 895 LIB148-049- LIB148 g596022 BLASTN 2194 1e−178 97 Q1-E1-D6 336 895 LIB3066-052- LIB3066 g596022 BLASTN 2178 1e−172 97 Q1-K1-G8 337 895 LIB148-016- LIB148 g596022 BLASTN 1567 1e−161 99 Q1-E1-G5 338 895 LIB143-032- LIB143 g596022 BLASTN 1914 1e−155 99 Q1-E1-E10 339 895 LIB3061-013- LIB3061 g596022 BLASTN 1738 1e−136 88 Q1-K1-F7 340 895 LIB143-047- LIB143 g596022 BLASTN 1490 1e−119 88 Q1-E1-D4 MAIZE VACUOLAR H+-TRANSLOCATING-PYROPHOSPHATASE 341 −700163331 700163331H1 SATMON013 g534915 BLASTN 751 1e−53 77 342 −700171438 700171438H1 SATMON013 g2258073 BLASTN 256 1e−10 76 343 −700202576 700202576H1 SATMON003 g2668746 BLASTX 214 1e−23 84 344 −700206487 700206487H1 SATMON003 g2570501 BLASTX 174 1e−17 86 345 −700217292 700217292H1 SATMON016 g2668746 BLASTX 214 1e−23 100 346 −700240889 700240889H1 SATMON010 g2570500 BLASTN 639 1e−47 84 347 −700347658 700347658H1 SATMON023 g2668746 BLASTX 215 1e−23 95 348 −700454151 700454151H1 SATMON029 g2668745 BLASTN 172 1e−10 90 349 −700454532 700454532H1 SATMON029 g2668745 BLASTN 259 1e−38 93 350 −700552133 700552133H1 SATMON022 g457744 BLASTX 176 1e−19 68 351 −700611864 700611864H1 SATMON022 g2668745 BLASTN 203 1e−9 84 352 107 700622451H1 SATMON034 g2668745 BLASTN 1645 1e−129 100 353 107 700571235H1 SATMON030 g2668745 BLASTN 1406 1e−125 98 354 107 700266126H1 SATMON017 g2668745 BLASTN 1145 1e−121 100 355 107 700621607H1 SATMON034 g2668745 BLASTN 1375 1e−121 99 356 107 700345080H1 SATMON021 g2668745 BLASTN 1195 1e−117 100 357 107 700624257H1 SATMON034 g2668745 BLASTN 825 1e−115 100 358 107 700030359H1 SATMON003 g2668745 BLASTN 1470 1e−114 100 359 107 700214462H1 SATMON016 g2668745 BLASTN 1223 1e−110 98 360 107 700356050H1 SATMON024 g2668745 BLASTN 1430 1e−110 100 361 107 701181128H1 SATMONN06 g2668745 BLASTN 1368 1e−105 98 362 107 700349795H1 SATMON023 g2668745 BLASTN 1370 1e−105 95 363 107 700473278H1 SATMON025 g2668745 BLASTN 1355 1e−104 100 364 107 700157057H1 SATMON012 g2668745 BLASTN 1345 1e−103 100 365 107 700622505H1 SATMON034 g2668745 BLASTN 762 1e−100 96 366 107 700219661H1 SATMON011 g2668745 BLASTN 942 1e−98 99 367 107 700619032H1 SATMON034 g2668745 BLASTN 989 1e−98 96 368 107 700620065H1 SATMON034 g2668745 BLASTN 1069 1e−98 94 369 107 700569179H1 SATMON030 g2668745 BLASTN 1233 1e−97 98 370 107 700156773H1 SATMON012 g2668745 BLASTN 1276 1e−97 99 371 107 700207120H1 SATMON017 g2668745 BLASTN 740 1e−96 99 372 107 700030407H1 SATMON003 g2668745 BLASTN 480 1e−95 98 373 107 700457309H1 SATMON029 g2668745 BLASTN 979 1e−95 99 374 107 700195681H1 SATMON014 g2668745 BLASTN 1246 1e−95 99 375 107 700444838H1 SATMON027 g2668745 BLASTN 1249 1e−95 96 376 107 700581619H1 SATMON031 g2668745 BLASTN 943 1e−94 96 377 107 700351021H1 SATMON023 g2668745 BLASTN 853 1e−91 92 378 107 700205723H1 SATMON003 g2668745 BLASTN 1138 1e−91 95 379 107 700159712H1 SATMON012 g2668745 BLASTN 1199 1e−91 94 380 107 700158937H1 SATMON012 g2668745 BLASTN 1132 1e−90 96 381 107 700336255H1 SATMON019 g2668745 BLASTN 489 1e−85 94 382 107 700422922H1 SATMONN01 g2668745 BLASTN 642 1e−84 95 383 107 700347429H1 SATMON023 g2668745 BLASTN 891 1e−83 92 384 107 700350695H1 SATMON023 g2668745 BLASTN 960 1e−83 91 385 107 700212988H1 SATMON016 g2668745 BLASTN 988 1e−82 96 386 107 700345278H1 SATMON021 g2668745 BLASTN 989 1e−82 95 387 107 700264475H1 SATMON017 g2668745 BLASTN 1089 1e−82 99 388 107 700211923H1 SATMON016 g2668745 BLASTN 991 1e−81 94 389 107 700620974H1 SATMON034 g2668745 BLASTN 907 1e−80 92 390 107 700156401H1 SATMON012 g2668745 BLASTN 1058 1e−79 90 391 107 700172547H1 SATMON013 g2668745 BLASTN 1042 1e−78 96 392 107 700552384H1 SATMON022 g2668745 BLASTN 916 1e−76 96 393 107 700219926H1 SATMON011 g2668745 BLASTN 1005 1e−75 100 394 107 700357492H1 SATMON024 g2668745 BLASTN 610 1e−74 99 395 107 700343365H1 SATMON021 g2668745 BLASTN 891 1e−74 94 396 107 700018618H1 SATMON001 g2668745 BLASTN 1001 1e−74 93 397 107 700570755H1 SATMON030 g2668745 BLASTN 845 1e−71 93 398 107 700194777H1 SATMON014 g2668745 BLASTN 940 1e−69 100 399 107 700453790H1 SATMON029 g2668745 BLASTN 925 1e−68 92 400 107 700197306H1 SATMON014 g2668745 BLASTN 928 1e−68 85 401 107 700355750H1 SATMON024 g2668745 BLASTN 393 1e−66 93 402 107 700172940H1 SATMON013 g2668745 BLASTN 902 1e−66 97 403 107 700102133H1 SATMON010 g2668745 BLASTN 850 1e−62 100 404 107 700350332H1 SATMON023 g2668745 BLASTN 539 1e−57 97 405 107 700450285H1 SATMON028 g2668745 BLASTN 750 1e−53 100 406 107 700165003H1 SATMON013 g2668745 BLASTN 548 1e−52 83 407 107 700016136H1 SATMON001 g2668745 BLASTN 527 1e−50 85 408 107 700171557H1 SATMON013 g2668745 BLASTN 714 1e−50 95 409 107 700238156H1 SATMON010 g2668745 BLASTN 715 1e−50 96 410 107 700425175H1 SATMONN01 g2668745 BLASTN 698 1e−49 94 411 107 700354402H1 SATMON024 g2668745 BLASTN 616 1e−48 91 412 107 700159204H1 SATMON012 g2668745 BLASTN 617 1e−42 94 413 107 700623602H1 SATMON034 g2668745 BLASTN 460 1e−38 100 414 107 700612844H1 SATMON033 g2668745 BLASTN 421 1e−36 84 415 107 700621062H2 SATMON034 g2668745 BLASTN 285 1e−25 89 416 107 700335685H1 SATMON019 g2668745 BLASTN 339 1e−25 91 417 13843 700334949H1 SATMON019 g2570500 BLASTN 680 1e−55 83 418 13843 700346817H1 SATMON021 g2570500 BLASTN 705 1e−54 83 419 13843 700103380H1 SATMON010 g2570500 BLASTN 710 1e−54 83 420 13843 700348280H1 SATMON023 g2570500 BLASTN 669 1e−51 83 421 13843 700453203H1 SATMON028 g2570500 BLASTN 659 1e−50 82 422 13843 700381101H1 SATMON023 g2570500 BLASTN 621 1e−47 82 423 13843 700347617H1 SATMON023 g2570500 BLASTN 592 1e−44 85 424 13843 700043259H1 SATMON004 g2570500 BLASTN 530 1e−39 84 425 13843 701184447H1 SATMONN06 g2570500 BLASTN 481 1e−35 78 426 21076 700241354H1 SATMON010 g166634 BLASTX 201 1e−20 58 427 24066 700423113H1 SATMONN01 g457744 BLASTX 124 1e−23 54 428 24266 700577157H1 SATMON031 g2570500 BLASTN 1001 1e−74 89 429 2531 700099364H1 SATMON009 g2570500 BLASTN 669 1e−51 86 430 2531 700336387H1 SATMON019 g2570500 BLASTN 389 1e−47 85 431 2531 700217095H1 SATMON016 g2570500 BLASTN 451 1e−33 86 432 2531 700155869H1 SATMON007 g2570500 BLASTN 385 1e−27 89 433 2531 700575534H1 SATMON030 g2570500 BLASTN 365 1e−26 88 434 2531 700163562H1 SATMON013 g2570501 BLASTX 145 1e−24 94 435 32364 700204306H1 SATMON003 g2668745 BLASTN 471 1e−28 74 436 32856 700166756H1 SATMON013 g534915 BLASTN 744 1e−53 76 437 32856 700042535H1 SATMON004 g534915 BLASTN 644 1e−44 73 438 3384 700237775H1 SATMON010 g2258073 BLASTN 911 1e−67 81 439 3384 700342456H1 SATMON021 g2258073 BLASTN 648 1e−64 78 440 3384 700073654H1 SATMON007 g2668745 BLASTN 860 1e−63 78 441 3384 700577805H1 SATMON031 g2258073 BLASTN 840 1e−61 78 442 3384 700028881H1 SATMON003 g534915 BLASTN 835 1e−60 78 443 3384 700215076H1 SATMON016 g534915 BLASTN 824 1e−59 78 444 3384 700017479H1 SATMON001 g534915 BLASTN 766 1e−55 80 445 3384 700204495H1 SATMON003 g534915 BLASTN 373 1e−51 81 446 3384 700206347H1 SATMON003 g2706449 BLASTN 685 1e−48 80 447 3384 700351040H1 SATMON023 g2706449 BLASTN 436 1e−45 78 448 3384 700345264H1 SATMON021 g2706449 BLASTN 616 1e−42 82 449 3384 700196795H1 SATMON014 g2570500 BLASTN 579 1e−39 80 450 3384 700019241H1 SATMON001 g2706449 BLASTN 583 1e−39 78 451 3384 700018612H1 SATMON001 g2668745 BLASTN 518 1e−34 76 452 3384 700102142H1 SATMON010 g2668745 BLASTN 539 1e−34 78 453 3384 700348430H1 SATMON023 g534915 BLASTN 489 1e−30 78 454 3384 700337745H1 SATMON020 g2706449 BLASTN 471 1e−28 79 455 3384 700439515H1 SATMON026 g534915 BLASTN 437 1e−27 75 456 3384 700074977H1 SATMON007 g534915 BLASTN 434 1e−25 76 457 3384 700615213H1 SATMON033 g2570501 BLASTX 125 1e−21 93 458 3384 700074109H1 SATMON007 g2668746 BLASTX 197 1e−20 72 459 3384 700549517H1 SATMON022 g2668746 BLASTX 172 1e−17 75 460 3384 700030347H1 SATMON003 g2668746 BLASTX 171 1e−16 77 461 3384 700221176H1 SATMON011 g2668746 BLASTX 171 1e−16 77 462 3384 700433360H1 SATMONN01 g2668746 BLASTX 95 1e−13 74 463 5000 700026151H1 SATMON003 g2903 BLASTX 261 1e−28 54 464 5000 700347165H1 SATMON021 g2624379 BLASTX 223 1e−24 51 465 5000 700430341H1 SATMONN01 g2903 BLASTX 185 1e−18 56 466 5000 700457781H1 SATMON029 g2903 BLASTX 133 1e−16 49 467 5861 700104993H1 SATMON010 g2258073 BLASTN 456 1e−27 73 468 5861 700203452H1 SATMON003 g2258073 BLASTN 428 1e−26 72 469 −L1431590 LIB143-006- LIB143 g16347 BLASTN 286 1e−13 61 Q1-E1-C9 470 −L1433414 LIB143-026- LIB143 g2258073 BLASTN 480 1e−29 70 Q1-E1-C3 471 −L1482832 LIB148-009- LIB148 g2258073 BLASTN 1086 1e−81 78 Q1-E1-D8 472 −L30674379 LIB3067-042- LIB3067 g2668745 BLASTN 305 1e−21 68 Q1-K1-H8 473 −L30675678 LIB3067-034- LIB3067 g2706449 BLASTN 286 1e−12 73 Q1-K1-E3 474 107 LIB3059-036- LIB3059 g2668745 BLASTN 1965 1e−166 100 Q1-K1-B10 475 107 LIB3061-035- LIB3061 g2668745 BLASTN 948 1e−138 93 Q1-K1-C9 476 107 LIB3061-032- LIB3061 g2668745 BLASTN 1685 1e−138 96 Q1-K1-A12 477 107 LIB3062-044- LIB3062 g2668745 BLASTN 1492 1e−134 95 Q1-K1-F8 478 107 LIB3068-025- LIB3068 g2668745 BLASTN 1687 1e−132 96 Q1-K1-E5 479 107 LIB3067-022- LIB3067 g2668745 BLASTN 1581 1e−128 91 Q1-K1-D11 480 107 LIB3067-016- LIB3067 g2668745 BLASTN 1305 1e−126 97 Q1-K1-G4 481 107 LIB3067-029- LIB3067 g2668745 BLASTN 1560 1e−125 90 Q1-K1-C6 482 107 LIB189-031- LIB189 g2668745 BLASTN 897 1e−81 85 Q1-E1-D3 483 24066 LIB3069-047- LIB3069 g166634 BLASTX 173 1e−45 55 Q1-K1-C4 484 24266 LIB3069-006- LIB3069 g2570500 BLASTN 717 1e−57 83 Q1-K1-F4 485 293 LIB3068-043- LIB3068 g633598 BLASTN 552 1e−34 78 Q1-K1-A2 486 32364 LIB3066-001- LIB3066 g2668745 BLASTN 612 1e−40 73 Q1-K1-B7 487 32856 LIB189-028- LIB189 g534915 BLASTN 986 1e−73 73 Q1-E1-C4 488 3384 LIB143-026- LIB143 g534915 BLASTN 1284 1e−98 78 Q1-E1-C1 489 3384 LIB3068-013- LIB3068 g534915 BLASTN 1074 1e−80 78 Q1-K1-H2 490 3384 LIB3062-033- LIB3062 g2668745 BLASTN 1009 1e−75 76 Q1-K1-D2 491 3384 LIB83-002- LIB83 g2706449 BLASTN 820 1e−59 78 Q1-E1-D2 492 3384 LIB3062-057- LIB3062 g2668745 BLASTN 801 1e−58 73 Q1-K1-B7 493 3384 LIB3062-001- LIB3062 g16347 BLASTN 802 1e−57 77 Q1-K2-H5 494 3384 LIB189-022- LIB189 g2668745 BLASTN 646 1e−43 75 Q1-E1-D5 495 3384 LIB189-012- LIB189 g2570501 BLASTX 138 1e−32 72 Q1-E1-F4 496 5000 LIB36-015- LIB36 g2624379 BLASTX 236 1e−41 51 Q1-E1-D6 497 5000 LIB83-016- LIB83 g4198 BLASTN 534 1e−33 61 Q1-E1-H7 MAIZE PYROPHOSPHATE-DEPENDENT FRUCTOSE-6-PHOSPHATE PHOSPHOTRANSFERASE 498 −700208959 700208959H1 SATMON016 g169538 BLASTX 107 1e−19 50 499 −700237606 700237606H1 SATMON010 g169538 BLASTX 114 1e−11 62 500 3456 700083478H1 SATMON011 g169538 BLASTX 121 1e−39 88 501 3652 700242182H1 SATMON010 g169538 BLASTX 155 1e−13 82 502 4965 700475352H1 SATMON025 g169538 BLASTX 123 1e−9 69 503 4965 700550752H1 SATMON022 g169538 BLASTX 123 1e−9 69 504 5359 700347441H1 SATMON023 g169538 BLASTX 139 1e−11 70 505 −L30594734 LIB3059-018- LIB3059 g169538 BLASTX 145 1e−49 83 Q1-K1-H3 506 −L30622375 LIB3062-009- LIB3062 g169538 BLASTX 157 1e−30 65 Q1-K1-B3 507 32156 L1B189-021- LIB189 g169538 BLASTX 123 1e−25 78 Q1-E1-G8 MAIZE INVERTASES 508 −700240132 700240132H1 SATMON010 g397631 BLASTX 134 1e−11 74 509 1923 700574932H1 SATMON030 g393390 BLASTX 152 1e−14 65 510 4355 700379641H1 SATMON021 g1177601 BLASTX 175 1e−19 85 MAIZE SUCROSE SYNTHASE 511 −700151470 700151470H1 SATMON007 g1196837 BLASTX 197 1e−27 64 512 −700214035 700214035H1 SATMON016 g22485 BLASTN 523 1e−34 79 513 −700262270 700262270H1 SATMON017 g2570066 BLASTN 866 1e−63 76 514 −700334686 700334686H1 SATMON019 g1100216 BLASTN 424 1e−31 88 515 −700381593 700381593H1 SATMON023 g22485 BLASTN 219 1e−13 97 516 −700404808 700404808H1 SATMON026 g2570066 BLASTN 859 1e−70 82 517 −700456905 700456905H1 SATMON029 g22485 BLASTN 528 1e−64 90 518 −700571529 700571529H1 SATMON030 g19106 BLASTX 139 1e−24 56 519 −700576567 700576567H1 SATMON030 g22485 BLASTN 285 1e−14 92 520 −700800659 700800659H1 SATMON036 g22485 BLASTN 558 1e−37 97 521 −700802941 700802941H1 SATMON036 g22485 BLASTN 316 1e−29 97 522 −701181030 701181030H1 SATMONN06 g2606080 BLASTN 669 1e−46 72 523 13723 700203023H1 SATMON003 g2570066 BLASTN 820 1e−68 84 524 13723 700215119H1 SATMON016 g2570066 BLASTN 680 1e−47 86 525 13723 700473266H1 SATMON025 g2570066 BLASTN 537 1e−35 85 526 15661 700440404H1 SATMON026 g2570066 BLASTN 364 1e−36 74 527 15661 700168252H1 SATMON013 g16525 BLASTN 433 1e−27 80 528 20925 700551647H1 SATMON022 g2570066 BLASTN 307 1e−35 73 529 20925 700257052H1 SATMON017 g2570067 BLASTX 118 1e−9 64 530 20934 700217752H1 SATMON016 g514945 BLASTN 1397 1e−107 98 531 20934 700332156H1 SATMON019 g514945 BLASTN 589 1e−97 95 532 30444 700257522H1 SATMON017 g1100216 BLASTN 760 1e−54 95 533 32909 700264718H1 SATMON017 g2570066 BLASTN 702 1e−57 76 534 405 700091402H1 SATMON011 g514945 BLASTN 1830 1e−143 100 535 405 700572549H1 SATMON030 g514945 BLASTN 1658 1e−129 99 536 405 700203058H1 SATMON003 g22485 BLASTN 1360 1e−127 100 537 405 700091753H1 SATMON011 g514945 BLASTN 1245 1e−126 99 538 405 700090929H1 SATMON011 g514945 BLASTN 1620 1e−126 100 539 405 700091711H1 SATMON011 g514945 BLASTN 1621 1e−126 99 540 405 700084254H1 SATMON011 g514945 BLASTN 1600 1e−124 100 541 405 700082305H1 SATMON011 g514945 BLASTN 1601 1e−124 99 542 405 700048236H1 SATMON003 g22485 BLASTN 1583 1e−123 99 543 405 700086713H1 SATMON011 g514945 BLASTN 1584 1e−123 99 544 405 700049353H1 SATMON003 g514945 BLASTN 1586 1e−123 99 545 405 700082766H1 SATMON011 g22485 BLASTN 1589 1e−123 98 546 405 700086055H1 SATMON011 g514945 BLASTN 1590 1e−123 100 547 405 700215105H1 SATMON016 g514945 BLASTN 1590 1e−123 100 548 405 700104149H1 SATMON010 g22485 BLASTN 1594 1e−123 98 549 405 700101601H1 SATMON009 g514945 BLASTN 1270 1e−122 100 550 405 700206869H1 SATMON003 g22485 BLASTN 1574 1e−122 97 551 405 700088163H1 SATMON011 g22485 BLASTN 1581 1e−122 99 552 405 700089166H1 SATMON011 g514945 BLASTN 1565 1e−121 100 553 405 700266251H1 SATMON017 g514945 BLASTN 1570 1e−121 100 554 405 700332710H1 SATMON019 g514945 BLASTN 1570 1e−121 100 555 405 700571106H1 SATMON030 g514945 BLASTN 1227 1e−120 98 556 405 700081893H1 SATMON011 g514945 BLASTN 1550 1e−120 98 557 405 700074739H1 SATMON007 g514945 BLASTN 1550 1e−120 100 558 405 700095163H1 SATMON008 g514945 BLASTN 1555 1e−120 100 559 405 700612766H1 SATMON033 g514945 BLASTN 883 1e−119 96 560 405 700267271H1 SATMON017 g514945 BLASTN 1535 1e−119 100 561 405 700083175H1 SATMON011 g514945 BLASTN 1535 1e−119 100 562 405 700088993H1 SATMON011 g22485 BLASTN 1545 1e−119 98 563 405 700094087H1 SATMON008 g22485 BLASTN 1526 1e−118 99 564 405 700086708H1 SATMON011 g514945 BLASTN 1529 1e−118 97 565 405 700090671H1 SATMON011 g514945 BLASTN 1530 1e−118 100 566 405 700209809H1 SATMON016 g22485 BLASTN 1532 1e−118 99 567 405 700084625H1 SATMON011 g514945 BLASTN 1533 1e−118 99 568 405 700089718H1 SATMON011 g514945 BLASTN 1120 1e−117 100 569 405 700213014H1 SATMON016 g514945 BLASTN 1405 1e−117 100 570 405 700086555H1 SATMON011 g514945 BLASTN 1514 1e−117 98 571 405 700475892H1 SATMON025 g514945 BLASTN 1516 1e−117 99 572 405 700047374H1 SATMON003 g22485 BLASTN 1516 1e−117 99 573 405 700090018H1 SATMON011 g514945 BLASTN 1519 1e−117 99 574 405 700076107H1 SATMON007 g514945 BLASTN 1520 1e−117 93 575 405 700213105H1 SATMON016 g514945 BLASTN 972 1e−116 99 576 405 700103806H1 SATMON010 g514945 BLASTN 1503 1e−116 99 577 405 700090748H1 SATMON011 g514945 BLASTN 1505 1e−116 100 578 405 700052006H1 SATMON003 g514945 BLASTN 1506 1e−116 99 579 405 700614963H1 SATMON033 g514945 BLASTN 957 1e−115 93 580 405 700337255H1 SATMON020 g22485 BLASTN 995 1e−115 97 581 405 700102778H1 SATMON010 g22485 BLASTN 1493 1e−115 99 582 405 700405466H1 SATMON029 g22485 BLASTN 1493 1e−115 99 583 405 700209634H1 SATMON016 g514945 BLASTN 1495 1e−115 100 584 405 700220467H1 SATMON011 g514945 BLASTN 1495 1e−115 100 585 405 700266637H1 SATMON017 g514945 BLASTN 1480 1e−114 100 586 405 700267579H1 SATMON017 g514945 BLASTN 1484 1e−114 99 587 405 700088475H1 SATMON011 g514945 BLASTN 1465 1e−113 100 588 405 700332618H1 SATMON019 g514945 BLASTN 1466 1e−113 99 589 405 700211347H1 SATMON016 g514945 BLASTN 1470 1e−113 100 590 405 700477206H1 SATMON025 g514945 BLASTN 1471 1e−113 99 591 405 700336768H1 SATMON019 g514945 BLASTN 1473 1e−113 99 592 405 700105305H1 SATMON010 g22485 BLASTN 1473 1e−113 99 593 405 700087114H1 SATMON011 g514945 BLASTN 1473 1e−113 99 594 405 700105366H1 SATMON010 g22485 BLASTN 1474 1e−113 98 595 405 700104831H1 SATMON010 g22485 BLASTN 825 1e−112 98 596 405 700620134H1 SATMON034 g22485 BLASTN 1179 1e−112 92 597 405 700211934H1 SATMON016 g22485 BLASTN 1215 1e−112 98 598 405 700096103H1 SATMON008 g514945 BLASTN 1391 1e−112 99 599 405 700264979H1 SATMON017 g514945 BLASTN 1454 1e−112 98 600 405 700053864H1 SATMON011 g514945 BLASTN 1455 1e−112 100 601 405 700211782H1 SATMON016 g514945 BLASTN 1460 1e−112 100 602 405 700102063H1 SATMON010 g22485 BLASTN 1461 1e−112 99 603 405 700207024H1 SATMON003 g514945 BLASTN 825 1e−111 100 604 405 700207970H1 SATMON016 g514945 BLASTN 1186 1e−111 98 605 405 700336624H1 SATMON019 g514945 BLASTN 1440 1e−111 100 606 405 700104357H1 SATMON010 g514945 BLASTN 1448 1e−111 98 607 405 700222053H1 SATMON011 g514945 BLASTN 1449 1e−111 99 608 405 700350806H1 SATMON023 g514945 BLASTN 660 1e−110 99 609 405 700091159H1 SATMON011 g514945 BLASTN 870 1e−110 100 610 405 700081810H1 SATMON011 g514945 BLASTN 926 1e−110 99 611 405 700102954H1 SATMON010 g514945 BLASTN 926 1e−110 97 612 405 700085307H1 SATMON011 g514945 BLASTN 1035 1e−110 100 613 405 700094295H1 SATMON008 g22485 BLASTN 1137 1e−110 96 614 405 700089176H1 SATMON011 g514945 BLASTN 1393 1e−110 97 615 405 700093643H1 SATMON008 g514945 BLASTN 1427 1e−110 95 616 405 700082421H1 SATMON011 g514945 BLASTN 1430 1e−110 98 617 405 700211788H1 SATMON016 g514945 BLASTN 1431 1e−110 99 618 405 700026724H1 SATMON003 g514945 BLASTN 1433 1e−110 97 619 405 700085275H1 SATMON011 g514945 BLASTN 1435 1e−110 100 620 405 700472161H1 SATMON025 g514945 BLASTN 755 1e−109 99 621 405 700084926H1 SATMON011 g514945 BLASTN 825 1e−109 100 622 405 700084592H1 SATMON011 g514945 BLASTN 920 1e−109 100 623 405 700053811H1 SATMON011 g514945 BLASTN 1296 1e−109 96 624 405 700216963H1 SATMON016 g514945 BLASTN 1415 1e−109 100 625 405 700085273H1 SATMON011 g22485 BLASTN 1416 1e−109 98 626 405 700082127H1 SATMON011 g514945 BLASTN 1420 1e−109 100 627 405 700085731H1 SATMON011 g514945 BLASTN 1425 1e−109 100 628 405 700088595H1 SATMON011 g22485 BLASTN 1426 1e−109 99 629 405 700470903H1 SATMON025 g514945 BLASTN 1426 1e−109 99 630 405 700265288H1 SATMON017 g514945 BLASTN 1375 1e−108 100 631 405 700072245H1 SATMON007 g514945 BLASTN 1404 1e−108 99 632 405 700347692H1 SATMON023 g514945 BLASTN 1405 1e−108 98 633 405 700214447H1 SATMON016 g514945 BLASTN 1406 1e−108 99 634 405 700476252H1 SATMON025 g514945 BLASTN 1407 1e−108 99 635 405 700336746H1 SATMON019 g514945 BLASTN 1409 1e−108 99 636 405 700053833H1 SATMON011 g514945 BLASTN 1410 1e−108 100 637 405 700094342H1 SATMON008 g514945 BLASTN 1410 1e−108 100 638 405 700202813H1 SATMON003 g514945 BLASTN 1032 1e−107 97 639 405 700050589H1 SATMON003 g514945 BLASTN 1035 1e−107 100 640 405 700050011H1 SATMON003 g514945 BLASTN 1078 1e−107 99 641 405 700215426H1 SATMON016 g514945 BLASTN 1189 1e−107 96 642 405 700472461H1 SATMON025 g514945 BLASTN 1392 1e−107 99 643 405 700336684H1 SATMON019 g22485 BLASTN 1393 1e−107 98 644 405 700449826H2 SATMON028 g514945 BLASTN 1395 1e−107 100 645 405 700216443H1 SATMON016 g514945 BLASTN 1396 1e−107 99 646 405 700240793H1 SATMON010 g514945 BLASTN 1399 1e−107 98 647 405 700215985H1 SATMON016 g514945 BLASTN 1400 1e−107 100 648 405 700336740H1 SATMON019 g514945 BLASTN 915 1e−106 99 649 405 700047958H1 SATMON003 g514945 BLASTN 987 1e−106 96 650 405 700085447H1 SATMON011 g514945 BLASTN 1030 1e−106 100 651 405 700084978H1 SATMON011 g514945 BLASTN 1121 1e−106 91 652 405 700800439H1 SATMON036 g22485 BLASTN 1379 1e−106 99 653 405 700219631H1 SATMON011 g514945 BLASTN 1380 1e−106 100 654 405 700220740H1 SATMON011 g514945 BLASTN 1380 1e−106 100 655 405 700243367H1 SATMON010 g514945 BLASTN 1381 1e−106 99 656 405 700220363H1 SATMON011 g514945 BLASTN 1387 1e−106 99 657 405 700215869H1 SATMON016 g514945 BLASTN 1390 1e−106 100 658 405 700216519H1 SATMON016 g514945 BLASTN 1131 1e−105 97 659 405 700052206H1 SATMON003 g514945 BLASTN 1264 1e−105 96 660 405 700094975H1 SATMON008 g514945 BLASTN 1368 1e−105 99 661 405 700220837H1 SATMON011 g514945 BLASTN 1369 1e−105 98 662 405 700221108H1 SATMON011 g514945 BLASTN 1370 1e−105 98 663 405 700222850H1 SATMON011 g514945 BLASTN 1370 1e−105 100 664 405 700214429H1 SATMON016 g514945 BLASTN 1373 1e−105 99 665 405 700473857H1 SATMON025 g514945 BLASTN 1375 1e−105 98 666 405 700213762H1 SATMON016 g514945 BLASTN 1378 1e−105 99 667 405 700405254H1 SATMON028 g22485 BLASTN 1242 1e−104 99 668 405 700029978H1 SATMON003 g22485 BLASTN 1324 1e−104 97 669 405 700238315H1 SATMON010 g514945 BLASTN 1355 1e−104 100 670 405 700241686H1 SATMON010 g514945 BLASTN 1358 1e−104 99 671 405 700237721H1 SATMON010 g22485 BLASTN 1360 1e−104 100 672 405 700217344H1 SATMON016 g514945 BLASTN 1360 1e−104 100 673 405 700030048H1 SATMON003 g514945 BLASTN 1363 1e−104 99 674 405 700211866H1 SATMON016 g514945 BLASTN 1363 1e−104 99 675 405 700214860H1 SATMON016 g514945 BLASTN 1365 1e−104 100 676 405 700085490H1 SATMON011 g514945 BLASTN 900 1e−103 98 677 405 700048568H1 SATMON003 g514945 BLASTN 980 1e−103 100 678 405 700381034H1 SATMON023 g22485 BLASTN 1269 1e−103 98 679 405 700220930H1 SATMON011 g514945 BLASTN 1347 1e−103 99 680 405 700030261H1 SATMON003 g514945 BLASTN 1353 1e−103 98 681 405 700081835H1 SATMON011 g22485 BLASTN 797 1e−102 98 682 405 700205270H1 SATMON003 g514945 BLASTN 1024 1e−102 94 683 405 700093612H1 SATMON008 g514945 BLASTN 1065 1e−102 99 684 405 700333392H1 SATMON019 g514945 BLASTN 1108 1e−102 97 685 405 700575385H1 SATMON030 g514945 BLASTN 1171 1e−102 96 686 405 700241061H1 SATMON010 g514945 BLASTN 1174 1e−102 99 687 405 700239916H1 SATMON010 g514945 BLASTN 1255 1e−102 100 688 405 700090248H1 SATMON011 g514945 BLASTN 1334 1e−102 98 689 405 700222923H1 SATMON011 g514945 BLASTN 1334 1e−102 98 690 405 700216993H1 SATMON016 g514945 BLASTN 1335 1e−102 100 691 405 700215984H1 SATMON016 g514945 BLASTN 1340 1e−102 100 692 405 700213182H1 SATMON016 g514945 BLASTN 1340 1e−102 98 693 405 700219845H1 SATMON011 g514945 BLASTN 1340 1e−102 100 694 405 700237762H1 SATMON010 g514945 BLASTN 1340 1e−102 100 695 405 700551043H1 SATMON022 g514945 BLASTN 1342 1e−102 99 696 405 700219254H1 SATMON011 g514945 BLASTN 1252 1e−101 99 697 405 700210348H1 SATMON016 g514945 BLASTN 1320 1e−101 97 698 405 700215089H1 SATMON016 g514945 BLASTN 1320 1e−101 100 699 405 700217251H1 SATMON016 g514945 BLASTN 1320 1e−101 100 700 405 700219240H1 SATMON011 g514945 BLASTN 1320 1e−101 100 701 405 700082094H1 SATMON011 g514945 BLASTN 1321 1e−101 99 702 405 700219385H1 SATMON011 g514945 BLASTN 1322 1e−101 99 703 405 700220052H1 SATMON011 g514945 BLASTN 1325 1e−101 100 704 405 700210366H1 SATMON016 g514945 BLASTN 1329 1e−101 93 705 405 700083089H1 SATMON011 g514945 BLASTN 1330 1e−101 98 706 405 700340286H1 SATMON020 g22485 BLASTN 677 1e−100 98 707 405 700221062H1 SATMON011 g514945 BLASTN 845 1e−100 98 708 405 700382272H1 SATMON024 g22485 BLASTN 958 1e−100 96 709 405 700209310H1 SATMON016 g514945 BLASTN 1187 1e−100 97 710 405 700052340H1 SATMON003 g514945 BLASTN 1188 1e−100 94 711 405 700467851H1 SATMON025 g22485 BLASTN 1245 1e−100 97 712 405 700088014H1 SATMON011 g514945 BLASTN 1270 1e−100 98 713 405 700214596H1 SATMON016 g514945 BLASTN 1295 1e−100 100 714 405 700157215H1 SATMON012 g22485 BLASTN 1310 1e−100 98 715 405 700223892H1 SATMON011 g514945 BLASTN 1310 1e−100 100 716 405 700218981H1 SATMON011 g514945 BLASTN 1310 1e−100 100 717 405 700081945H1 SATMON011 g514945 BLASTN 1311 1e−100 96 718 405 700217817H1 SATMON016 g514945 BLASTN 1315 1e−100 100 719 405 700469042H1 SATMON025 g514945 BLASTN 561 1e−99 98 720 405 700474709H1 SATMON025 g514945 BLASTN 801 1e−99 99 721 405 700201736H1 SATMON003 g514945 BLASTN 1168 1e−99 98 722 405 700223516H1 SATMON011 g514945 BLASTN 1201 1e−99 99 723 405 700453941H1 SATMON029 g22485 BLASTN 1295 1e−99 95 724 405 700212970H1 SATMON016 g514945 BLASTN 1297 1e−99 99 725 405 700215662H1 SATMON016 g22485 BLASTN 1297 1e−99 99 726 405 700802209H1 SATMON036 g22485 BLASTN 1300 1e−99 98 727 405 700343716H1 SATMON021 g514945 BLASTN 1300 1e−99 100 728 405 700223322H1 SATMON011 g514945 BLASTN 1300 1e−99 100 729 405 700217238H1 SATMON016 g514945 BLASTN 1300 1e−99 100 730 405 700195066H1 SATMON014 g22485 BLASTN 1300 1e−99 98 731 405 700072395H1 SATMON007 g514945 BLASTN 1301 1e−99 95 732 405 700212752H1 SATMON016 g22485 BLASTN 1305 1e−99 98 733 405 700222204H1 SATMON011 g514945 BLASTN 1305 1e−99 100 734 405 700550572H1 SATMON022 g22485 BLASTN 713 1e−98 97 735 405 700213879H1 SATMON016 g514945 BLASTN 866 1e−98 99 736 405 700551585H1 SATMON022 g514945 BLASTN 916 1e−98 99 737 405 700195025H1 SATMON014 g22485 BLASTN 1283 1e−98 98 738 405 700800710H1 SATMON036 g22485 BLASTN 1283 1e−98 98 739 405 700222985H1 SATMON011 g22485 BLASTN 1283 1e−98 98 740 405 700798823H1 SATMON036 g22485 BLASTN 1284 1e−98 98 741 405 700104391H1 SATMON010 g22485 BLASTN 1289 1e−98 98 742 405 700466592H1 SATMON025 g22485 BLASTN 1289 1e−98 95 743 405 700027037H1 SATMON003 g514945 BLASTN 919 1e−97 91 744 405 700214371H1 SATMON016 g514945 BLASTN 1033 1e−97 95 745 405 700799077H1 SATMON036 g22485 BLASTN 1091 1e−97 99 746 405 700467028H1 SATMON025 g514945 BLASTN 1103 1e−97 98 747 405 700219393H1 SATMON011 g514945 BLASTN 1250 1e−97 99 748 405 700197602H1 SATMON014 g22485 BLASTN 1273 1e−97 97 749 405 700801226H1 SATMON036 g22485 BLASTN 1276 1e−97 99 750 405 700216371H1 SATMON016 g514945 BLASTN 1278 1e−97 98 751 405 700805695H1 SATMON036 g22485 BLASTN 1280 1e−97 98 752 405 700334076H1 SATMON019 g514945 BLASTN 503 1e−96 98 753 405 700082647H1 SATMON011 g514945 BLASTN 735 1e−96 100 754 405 700458687H1 SATMON029 g22485 BLASTN 751 1e−96 95 755 405 700220750H1 SATMON011 g514945 BLASTN 1187 1e−96 96 756 405 700194931H1 SATMON014 g22485 BLASTN 1262 1e−96 99 757 405 700800522H1 SATMON036 g22485 BLASTN 1265 1e−96 98 758 405 700244185H1 SATMON010 g514945 BLASTN 1265 1e−96 100 759 405 700240785H1 SATMON010 g514945 BLASTN 1268 1e−96 98 760 405 700551959H1 SATMON022 g514945 BLASTN 1270 1e−96 100 761 405 700085057H1 SATMON011 g514945 BLASTN 682 1e−95 97 762 405 700332020H1 SATMON019 g514945 BLASTN 713 1e−95 97 763 405 700208841H1 SATMON016 g514945 BLASTN 822 1e−95 95 764 405 700193023H1 SATMON014 g22485 BLASTN 1248 1e−95 98 765 405 700153902H1 SATMON007 g514945 BLASTN 1250 1e−95 100 766 405 700196173H1 SATMON014 g22485 BLASTN 1252 1e−95 99 767 405 700804187H1 SATMON036 g22485 BLASTN 1253 1e−95 98 768 405 700339656H1 SATMON020 g22485 BLASTN 1257 1e−95 99 769 405 700238537H1 SATMON010 g22485 BLASTN 1258 1e−95 99 770 405 700551221H1 SATMON022 g514945 BLASTN 1204 1e−94 99 771 405 700801807H1 SATMON036 g22485 BLASTN 1235 1e−94 100 772 405 700085096H1 SATMON011 g514945 BLASTN 1235 1e−94 100 773 405 700020516H1 SATMON001 g514945 BLASTN 1236 1e−94 98 774 405 700193482H1 SATMON014 g22485 BLASTN 1236 1e−94 99 775 405 700217793H1 SATMON016 g514945 BLASTN 1237 1e−94 98 776 405 700088752H1 SATMON011 g514945 BLASTN 1240 1e−94 100 777 405 700087940H1 SATMON011 g514945 BLASTN 1241 1e−94 99 778 405 700089541H1 SATMON011 g533251 BLASTN 1067 1e−93 91 779 405 700346461H1 SATMON021 g514945 BLASTN 1190 1e−93 99 780 405 700156258H1 SATMON007 g514945 BLASTN 1225 1e−93 100 781 405 700195532H1 SATMON014 g22485 BLASTN 1226 1e−93 99 782 405 700196548H1 SATMON014 g22485 BLASTN 1227 1e−93 99 783 405 700213282H1 SATMON016 g514945 BLASTN 1227 1e−93 96 784 405 700194939H1 SATMON014 g22485 BLASTN 1228 1e−93 98 785 405 700340787H1 SATMON020 g22485 BLASTN 697 1e−92 94 786 405 700105064H1 SATMON010 g22485 BLASTN 706 1e−92 98 787 405 700805132H1 SATMON036 g22485 BLASTN 1046 1e−92 99 788 405 700803414H1 SATMON036 g22485 BLASTN 1211 1e−92 99 789 405 700214594H1 SATMON016 g514945 BLASTN 1212 1e−92 98 790 405 700215080H1 SATMON016 g514945 BLASTN 1213 1e−92 92 791 405 700224219H1 SATMON011 g514945 BLASTN 1213 1e−92 99 792 405 700048190H1 SATMON003 g514945 BLASTN 1218 1e−92 92 793 405 700223284H1 SATMON011 g51494S BLASTN 1220 1e−92 100 794 405 700152413H1 SATMON007 g514945 BLASTN 1220 1e−92 100 795 405 700218003H1 SATMON016 g514945 BLASTN 1221 1e−92 96 796 405 700160155H1 SATMON012 g22485 BLASTN 1222 1e−92 99 797 405 700087943H1 SATMON011 g22485 BLASTN 1222 1e−92 99 798 405 700474049H1 SATMON025 g514945 BLASTN 632 1e−91 97 799 405 700216994H1 SATMON016 g514945 BLASTN 1043 1e−91 99 800 405 700346892H1 SATMON021 g514945 BLASTN 1210 1e−91 96 801 405 700142782H1 SATMON013 g514945 BLASTN 1190 1e−90 100 802 405 700244157H1 SATMON010 g514945 BLASTN 1197 1e−90 97 803 405 700469243H1 SATMON025 g22485 BLASTN 701 1e−89 98 804 405 700618387H1 SATMON033 g514945 BLASTN 853 1e−89 93 805 405 700211154H1 SATMON016 g5332S1 BLASTN 917 1e−89 90 806 405 700081933H1 SATMON011 g533251 BLASTN 955 1e−89 91 807 405 700235229H1 SATMON010 g514945 BLASTN 955 1e−89 97 808 405 700209241H1 SATMON016 g514945 BLASTN 1076 1e−89 98 809 405 700265841H1 SATMON017 g514945 BLASTN 1097 1e−89 95 810 405 700193804H1 SATMON014 g22485 BLASTN 1180 1e−89 98 811 405 700167742H1 SATMON013 g22485 BLASTN 1180 1e−89 100 812 405 700163256H1 SATMON013 g514945 BLASTN 1182 1e−89 97 813 405 700184973H1 SATMON014 g22485 BLASTN 1182 1e−89 99 814 405 700171785H1 SATMON013 g22485 BLASTN 1184 1e−89 97 815 405 700216915H1 SATMON016 g514945 BLASTN 1185 1e−89 100 816 405 700806685H1 SATMON036 g22485 BLASTN 1186 1e−89 99 817 405 700803846H1 SATMON036 g22485 BLASTN 892 1e−88 95 818 405 700218514H1 SATMON011 g533251 BLASTN 907 1e−88 91 819 405 700574674H1 SATMON030 g22485 BLASTN 957 1e−88 84 820 405 700196082H1 SATMON014 g22485 BLASTN 1054 1e−88 94 821 405 700241637H1 SATMON010 g22485 BLASTN 1081 1e−88 98 822 405 700801876H1 SATMON036 g22485 BLASTN 1163 1e−88 98 823 405 700213443H1 SATMON016 g514945 BLASTN 1171 1e−88 99 824 405 700465181H1 SATMON025 g22485 BLASTN 797 1e−87 92 825 405 700798732H1 SATMON036 g22485 BLASTN 1159 1e−87 96 826 405 700153473H1 SATMON007 g514945 BLASTN 1160 1e−87 100 827 405 700165496H1 SATMON013 g514945 BLASTN 1160 1e−87 97 828 405 700264250H1 SATMON017 g514945 BLASTN 643 1e−86 100 829 405 700335490H1 SATMON019 g514945 BLASTN 671 1e−86 97 830 405 700575891H1 SATMON030 g22485 BLASTN 780 1e−86 92 831 405 700222931H1 SATMON011 g514945 BLASTN 1117 1e−86 91 832 405 700163588H1 SATMON013 g514945 BLASTN 1140 1e−86 100 833 405 700161111H1 SATMON012 g22485 BLASTN 1142 1e−86 99 834 405 700016023H1 SATMON001 g514945 BLASTN 1147 1e−86 99 835 405 700209043H1 SATMON016 g514945 BLASTN 659 1e−85 98 836 405 700333556H1 SATMON019 g514945 BLASTN 783 1e−85 89 837 405 700570471H1 SATMON030 g22485 BLASTN 831 1e−85 89 838 405 700171040H1 SATMON013 g22485 BLASTN 1127 1e−85 99 839 405 700196146H1 SATMON014 g22485 BLASTN 1128 1e−85 96 840 405 700021837H1 SATMON001 g514945 BLASTN 1130 1e−85 98 841 405 700169062H1 SATMON013 g514945 BLASTN 1130 1e−85 100 842 405 700218263H1 SATMON016 g514945 BLASTN 1135 1e−85 100 843 405 700091281H1 SATMON011 g514945 BLASTN 1135 1e−85 100 844 405 700221221H1 SATMON011 g22485 BLASTN 735 1e−84 98 845 405 700239933H1 SATMON010 g22485 BLASTN 779 1e−84 97 846 405 700193089H1 SATMON014 g22485 BLASTN 979 1e−84 97 847 405 700216767H1 SATMON016 g514945 BLASTN 1005 1e−84 98 848 405 700085786H1 SATMON011 g514945 BLASTN 1117 1e−84 99 849 405 700163868H1 SATMON013 g22485 BLASTN 1120 1e−84 100 850 405 700167028H1 SATMON013 g514945 BLASTN 1120 1e−84 100 851 405 700170416H1 SATMON013 g514945 BLASTN 1120 1e−84 100 852 405 700266595H1 SATMON017 g514945 BLASTN 1120 1e−84 100 853 405 700377630H1 SATMON019 g514945 BLASTN 649 1e−83 95 854 405 700207830H1 SATMON016 g514945 BLASTN 877 1e−83 97 855 405 700241726H1 SATMON010 g514945 BLASTN 1104 1e−83 97 856 405 700806447H1 SATMON036 g22485 BLASTN 1106 1e−83 93 857 405 700018166H1 SATMON001 g514945 BLASTN 1108 1e−83 98 858 405 700083463H1 SATMON011 g514945 BLASTN 633 1e−82 92 859 405 700548890H1 SATMON022 g22485 BLASTN 727 1e−82 93 860 405 700218553H1 SATMON011 g22485 BLASTN 979 1e−82 95 861 405 700016408H1 SATMON001 g514945 BLASTN 1026 1e−82 97 862 405 700569427H2 SATMON030 g514945 BLASTN 1095 1e−82 97 863 405 700172546H1 SATMON013 g514945 BLASTN 1100 1e−82 100 864 405 700804387H1 SATMON036 g22485 BLASTN 668 1e−81 97 865 405 700155788H1 SATMON007 g514945 BLASTN 840 1e−81 100 866 405 700807167H1 SATMON036 g22485 BLASTN 1024 1e−81 97 867 405 700472356H1 SATMON025 g22485 BLASTN 1080 1e−81 98 868 405 700193535H1 SATMON014 g22485 BLASTN 1082 1e−81 99 869 405 700171177H1 SATMON013 g22485 BLASTN 1086 1e−81 98 870 405 700799867H1 SATMON036 g22485 BLASTN 1087 1e−81 96 871 405 700263716H1 SATMON017 g514945 BLASTN 1089 1e−81 92 872 405 700476045H1 SATMON025 g22485 BLASTN 608 1e−80 88 873 405 700803344H1 SATMON036 g22485 BLASTN 834 1e−80 97 874 405 700168924H1 SATMON013 g514945 BLASTN 860 1e−80 99 875 405 700218569H1 SATMON011 g22485 BLASTN 900 1e−80 98 876 405 700088574H1 SATMON011 g514945 BLASTN 900 1e−80 86 877 405 700471932H1 SATMON025 g530978 BLASTN 1064 1e−80 83 878 405 700020011H1 SATMON001 g22485 BLASTN 1067 1e−80 99 879 405 700167511H1 SATMON013 g22485 BLASTN 1070 1e−80 100 880 405 700219249H1 SATMON011 g514945 BLASTN 1070 1e−80 100 881 405 700804846H1 SATMON036 g22485 BLASTN 1075 1e−80 90 882 405 700150388H1 SATMON007 g22485 BLASTN 1075 1e−80 100 883 405 700807395H1 SATMON036 g22485 BLASTN 571 1e−79 90 884 405 700090864H1 SATMON011 g514945 BLASTN 630 1e−79 100 885 405 700217812H1 SATMON016 g514945 BLASTN 646 1e−79 91 886 405 700203618H1 SATMON003 g22485 BLASTN 913 1e−79 96 887 405 700203302H1 SATMON003 g514945 BLASTN 1030 1e−79 100 888 405 700163192H1 SATMON013 g22485 BLASTN 1056 1e−79 97 889 405 700805065H1 SATMON036 g22485 BLASTN 1066 1e−79 95 890 405 700086763H1 SATMON011 g514945 BLASTN 901 1e−78 98 891 405 700240070H1 SATMON010 g533251 BLASTN 923 1e−78 90 892 405 700018847H1 SATMON001 g22485 BLASTN 1045 1e−78 98 893 405 700803420H1 SATMON036 g22485 BLASTN 1048 1e−78 96 894 405 700799936H1 SATMON036 g22485 BLASTN 1050 1e−78 96 895 405 700207637H1 SATMON016 g514945 BLASTN 828 1e−77 98 896 405 700807034H1 SATMON036 g22485 BLASTN 808 1e−76 91 897 405 700198035H1 SATMON016 g514945 BLASTN 1025 1e−76 100 898 405 700169076H1 SATMON013 g514945 BLASTN 1028 1e−76 99 899 405 700020564H1 SATMON001 g514945 BLASTN 1030 1e−76 98 900 405 700799968H1 SATMON036 g22485 BLASTN 701 1e−75 99 901 405 700378056H1 SATMON019 g22485 BLASTN 802 1e−75 97 902 405 700219691H1 SATMON011 g514945 BLASTN 1018 1e−75 99 903 405 700168945H1 SATMON013 g22485 BLASTN 848 1e−74 94 904 405 700242730H1 SATMON010 g514945 BLASTN 1006 1e−74 99 905 405 700210096H1 SATMON016 g514945 BLASTN 756 1e−73 93 906 405 700333941H1 SATMON019 g514945 BLASTN 923 1e−73 99 907 405 700576645H1 SATMON030 g22485 BLASTN 991 1e−73 99 908 405 700333494H1 SATMON019 g514945 BLASTN 601 1e−72 91 909 405 700023296H1 SATMON003 g514945 BLASTN 726 1e−72 95 910 405 700802508H1 SATMON036 g22485 BLASTN 811 1e−72 94 911 405 700223382H1 SATMON011 g22485 BLASTN 865 1e−72 98 912 405 700215535H1 SATMON016 g514945 BLASTN 942 1e−72 96 913 405 700017549H1 SATMON001 g514945 BLASTN 973 1e−72 97 914 405 700799113H1 SATMON036 g22485 BLASTN 787 1e−70 99 915 405 700168696H1 SATMON013 g514945 BLASTN 946 1e−69 89 916 405 700088269H1 SATMON011 g514945 BLASTN 946 1e−69 93 917 405 700194522H1 SATMON014 g22485 BLASTN 875 1e−68 97 918 405 700203476H1 SATMON003 g22485 BLASTN 923 1e−68 86 919 405 700549205H1 SATMON022 g22485 BLASTN 300 1e−66 89 920 405 700196217H1 SATMON014 g22485 BLASTN 907 1e−66 96 921 405 700163647H1 SATMON013 g22485 BLASTN 888 1e−65 98 922 405 700804485H1 SATMON036 g22485 BLASTN 896 1e−65 99 923 405 700193074H1 SATMON014 g22485 BLASTN 605 1e−63 96 924 405 700203370H1 SATMON003 g514945 BLASTN 857 1e−62 98 925 405 700201575H1 SATMON003 g514945 BLASTN 335 1e−60 87 926 405 700378020H1 SATMON019 g514945 BLASTN 833 1e−60 97 927 405 700242865H1 SATMON010 g514945 BLASTN 823 1e−59 91 928 405 700344036H1 SATMON021 g514945 BLASTN 825 1e−59 100 929 405 700215849H1 SATMON016 g514945 BLASTN 805 1e−58 100 930 405 700443538H1 SATMON027 g22485 BLASTN 814 1e−58 98 931 405 700804448H1 SATMON036 g22485 BLASTN 791 1e−57 99 932 405 700155008H1 SATMON007 g22485 BLASTN 802 1e−57 98 933 405 700201244H1 SATMON003 g22485 BLASTN 530 1e−56 97 934 405 700616378H1 SATMON033 g22485 BLASTN 682 1e−56 97 935 405 700333357H1 SATMON019 g22485 BLASTN 780 1e−56 80 936 405 700222360H1 SATMON011 g514945 BLASTN 777 1e−55 92 937 405 700214724H1 SATMON016 g514945 BLASTN 763 1e−54 98 938 405 700571283H1 SATMON030 g514945 BLASTN 736 1e−52 99 939 405 700020194H1 SATMON001 g22485 BLASTN 415 1e−51 99 940 405 700620551H1 SATMON034 g22485 BLASTN 473 1e−51 95 941 405 700446320H1 SATMON027 g22485 BLASTN 475 1e−50 87 942 405 700241357H1 SATMON010 g22485 BLASTN 701 1e−49 99 943 405 700617094H1 SATMON033 g22485 BLASTN 673 1e−47 97 944 405 700206691H1 SATMON003 g514945 BLASTN 680 1e−47 90 945 405 700091580H1 SATMON011 g514945 BLASTN 680 1e−47 100 946 405 700574515H1 SATMON030 g514945 BLASTN 369 1e−46 74 947 405 700155148H1 SATMON007 g514945 BLASTN 397 1e−45 97 948 405 700612388H1 SATMON033 g514945 BLASTN 625 1e−43 100 949 405 700474681H1 SATMON025 g22485 BLASTN 379 1e−41 91 950 405 700800401H1 SATMON036 g22485 BLASTN 395 1e−40 90 951 405 700155657H1 SATMON007 g514945 BLASTN 591 1e−40 95 952 405 700076002H1 SATMON007 g514945 BLASTN 575 1e−39 100 953 405 700802090H1 SATMON036 g22485 BLASTN 577 1e−39 98 954 405 700170104H1 SATMON013 g22485 BLASTN 565 1e−38 100 955 405 701183763H1 SATMONN06 g514945 BLASTN 569 1e−38 90 956 405 700084688H1 SATMON011 g514945 BLASTN 380 1e−36 98 957 405 700473655H1 SATMON025 g22485 BLASTN 530 1e−35 100 958 405 700615166H1 SATMON033 g514945 BLASTN 531 1e−35 94 959 405 700085562H1 SATMON011 g533251 BLASTN 532 1e−35 98 960 405 700153049H1 SATMON007 g514945 BLASTN 537 1e−35 94 961 405 700090656H1 SATMON011 g514945 BLASTN 489 1e−34 98 962 405 700802054H1 SATMON036 g22485 BLASTN 345 1e−31 99 963 405 700802284H1 SATMON036 g22485 BLASTN 488 1e−31 97 964 405 700802312H1 SATMON036 g22485 BLASTN 270 1e−30 100 965 405 700153683H1 SATMON007 g22485 BLASTN 461 1e−29 98 966 405 700028453H1 SATMON003 g22485 BLASTN 321 1e−27 99 967 405 700089391H1 SATMON011 g514945 BLASTN 404 1e−24 96 968 405 700381969H1 SATMON023 g22485 BLASTN 385 1e−23 94 969 405 700800135H1 SATMON036 g22485 BLASTN 180 1e−21 100 970 405 700088173H1 SATMON011 g514945 BLASTN 347 1e−20 95 971 405 700202170H1 SATMON003 g19108 BLASTX 133 1e−11 96 972 537 700209929H1 SATMON016 g22485 BLASTN 1478 1e−114 99 973 537 700096948H1 SATMON008 g22485 BLASTN 911 1e−113 99 974 537 700476287H1 SATMON025 g22485 BLASTN 1403 1e−108 98 975 537 700803088H1 SATMON036 g22485 BLASTN 1336 1e−107 96 976 537 700799436H1 SATMON036 g22485 BLASTN 1361 1e−104 99 977 537 700224822H1 SATMON011 g22485 BLASTN 1300 1e−103 96 978 537 700241134H1 SATMON010 g22485 BLASTN 1302 1e−99 99 979 537 700803625H1 SATMON036 g22485 BLASTN 1292 1e−98 99 980 537 700802549H1 SATMON036 g22485 BLASTN 1232 1e−93 99 981 537 700477992H1 SATMON025 g22485 BLASTN 943 1e−92 97 982 537 700150953H1 SATMON007 g22485 BLASTN 1152 1e−87 99 983 537 700205638H1 SATMON003 g22485 BLASTN 1086 1e−81 99 984 537 700803732H1 SATMON036 g22487 BLASTN 379 1e−79 97 985 537 700165461H1 SATMON013 g22485 BLASTN 1064 1e−79 98 986 537 700807069H1 SATMON036 g22485 BLASTN 957 1e−77 96 987 537 700800902H1 SATMON036 g22485 BLASTN 762 1e−54 86 988 537 700466671H1 SATMON025 g22485 BLASTN 520 1e−44 95 989 537 700799118H1 SATMON036 g22485 BLASTN 626 1e−43 99 990 537 700802273H1 SATMON036 g22485 BLASTN 616 1e−42 99 991 537 700804848H1 SATMON036 g22485 BLASTN 306 1e−33 98 992 8549 700103190H1 SATMON010 g1100216 BLASTN 615 1e−92 98 993 8549 700075574H1 SATMON007 g1100216 BLASTN 701 1e−92 100 994 8549 700218547H1 SATMON011 g514945 BLASTN 1208 1e−91 99 995 8549 700213873H1 SATMON016 g1100216 BLASTN 673 1e−90 95 996 8549 700221147H1 SATMON011 g1100216 BLASTN 646 1e−89 98 997 8549 700207093H1 SATMON003 g1100216 BLASTN 701 1e−87 100 998 8549 700210112H1 SATMON016 g1100216 BLASTN 615 1e−84 98 999 8549 700096984H1 SATMON008 g514945 BLASTN 1111 1e−83 99 1000 8549 700221070H1 SATMON011 g1100216 BLASTN 645 1e−82 96 1001 8549 700332046H1 SATMON019 g1100216 BLASTN 601 1e−76 89 1002 8549 700150377H1 SATMON007 g1100216 BLASTN 621 1e−74 100 1003 8549 700084780H1 SATMON011 g514945 BLASTN 585 1e−39 100 1004 8549 700153082H1 SATMON007 g1100216 BLASTN 495 1e−36 89 1005 8549 700261144H1 SATMON017 g1100216 BLASTN 339 1e−35 87 1006 8549 700264112H1 SATMON017 g1100216 BLASTN 428 1e−34 91 1007 8549 700473660H1 SATMON025 g1100216 BLASTN 415 1e−28 100 1008 8549 700473628H1 SATMON025 g514945 BLASTN 329 1e−26 88 1009 8549 700351060H1 SATMON023 g1100216 BLASTN 291 1e−22 91 1010 −L30595280 LIB3059-039- LIB3059 g22485 BLASTN 473 1e−30 79 Q1-K1-A5 1011 −L30612133 LIB3061-024- LIB3061 g22485 BLASTN 849 1e−61 80 Q1-K1-H5 1012 −L30616296 LIB3061-043- LIB3061 g22485 BLASTN 479 1e−98 82 Q1-K1-A10 1013 −L30623037 LIB3062-030- LIB3062 g514945 BLASTN 684 1e−48 78 Q1-K1-F12 1014 −L30625289 LIB3062-021- LIB3062 g514945 BLASTN 1180 1e−111 79 Q1-K1-C2 1015 −L30663565 LIB3066-053- LIB3066 g530978 BLASTN 568 1e−36 76 Q1-K1-D6 1016 −L30784420 LIB3078-039- LIB3078 g514945 BLASTN 484 1e−40 81 Q1-K1-A4 1017 30444 LIB3069-052- LIB3069 g1100216 BLASTN 558 1e−77 89 Q1-K1-F8 1018 32909 LIB143-057- LIB143 g2570066 BLASTN 902 1e−69 74 Q1-E1-F6 1019 405 LIB3062-021- LIB3062 g514945 BLASTN 2368 1e−188 99 Q1-K1-C5 1020 405 LIB3078-024- LIB3078 g514945 BLASTN 2356 1e−187 98 Q1-KI-C5 1021 405 LIB3059-028- LIB3059 g22485 BLASTN 2163 1e−171 98 Q1-KI-D5 1022 405 LIB3059-015- LIB3059 g22485 BLASTN 2167 1e−171 98 Q1-K1-E7 1023 405 LIB3059-044- LIB3059 g514945 BLASTN 2170 1e−171 98 Q1-K1-E7 1024 405 LIB3061-029- LIB3061 g22485 BLASTN 2055 1e−170 98 Q1-K1-G11 1025 405 LIB3059-011- LIB3059 g22485 BLASTN 2137 1e−169 98 Q1-K1-F5 1026 405 LIB3062-009- LIB3062 g514945 BLASTN 2122 1e−167 98 Q1-K1-D1 1027 405 LIB3061-011- LIB3061 g22485 BLASTN 2091 1e−165 98 Q1-K1-D9 1028 405 LIB3067-040- LIB3067 g514945 BLASTN 1916 1e−164 99 Q1-K1-E8 1029 405 LIB3062-041- LIB3062 g514945 BLASTN 2082 1e−164 97 Q1-K1-D4 1030 405 LIB3062-022- LIB3062 g514945 BLASTN 2084 1e−164 99 Q1-K1-C9 1031 405 LIB3062-033- LIB3062 g514945 BLASTN 1854 1e−161 95 Q1-KI-C7 1032 405 LIB3062-002- LIB3062 g514945 BLASTN 1854 1e−161 97 Q1-K2-F9 1033 405 LIB3059-010- LIB3059 g22485 BLASTN 2018 1e−159 99 Q1-K1-C9 1034 405 LIB3059-013- LIB3059 g22485 BLASTN 2022 1e−159 98 Q1-K1-B10 1035 405 LIB3061-020- LIB3061 g22485 BLASTN 1771 1e−158 97 Q1-K1-F2 1036 405 LIB3061-022- LIB3061 g22485 BLASTN 1909 1e−158 98 Q1-K1-C2 1037 405 LIB3062-023- LIB3062 g22485 BLASTN 1508 1e−157 96 Q1-K1-D10 1038 405 LIB3061-008- LIB3061 g22485 BLASTN 1983 1e−156 97 Q1-K1-H11 1039 405 LIB3059-024- LIB3059 g22485 BLASTN 1051 1e−154 99 Q1-K1-H4 1040 405 LIB3062-048- LIB3062 g22485 BLASTN 1187 1e−154 94 Q1-K1-G5 1041 405 LIB3061-025- LIB3061 g22485 BLASTN 1803 1e−154 95 Q1-K1-B1 1042 405 LIB3061-028- LIB3061 g22485 BLASTN 1963 1e−154 97 Q1-K1-C4 1043 405 LIB3078-057- LIB3078 g514945 BLASTN 1412 1e−153 92 Q1-K1-D9 1044 405 LIB3061-021- LIB3061 g22485 BLASTN 1465 1e−153 96 Q1-K1-A8 1045 405 LIB3061-025- LIB3061 g22485 BLASTN 1524 1e−153 96 Q1-K1-B5 1046 405 LIB3061-008- LIB3061 g22485 BLASTN 1879 1e−153 94 Q1-K1-C7 1047 405 LIB3078-039- LIB3078 g514945 BLASTN 1853 1e−151 96 Q1-K1-A8 1048 405 LIB3061-049- LIB3061 g22485 BLASTN 1801 1e−150 98 Q1-K1-E5 1049 405 LIB3062-001- LIB3062 g514945 BLASTN 1916 1e−150 94 Q1-K2-G2 1050 405 LIB3061-021- LIB3061 g22485 BLASTN 1918 1e−150 92 Q1-K1-G6 1051 405 LIB3061-039- LIB3061 g22485 BLASTN 1361 1e−149 96 Q1-K1-D2 1052 405 LIB3061-051- LIB3061 g22485 BLASTN 1768 1e−148 98 Q1-K1-G8 1053 405 LIB3061-015- LIB3061 g22485 BLASTN 1667 1e−146 93 Q1-K1-A12 1054 405 LIB3059-040- LIB3059 g22485 BLASTN 1835 1e−146 97 Q1-K1-H11 1055 405 LIB3061-002- LIB3061 g22485 BLASTN 1845 1e−144 89 Q1-K2-G5 1056 405 LIB3062-002- LIB3062 g22485 BLASTN 1672 1e−142 99 Q1-K2-G12 1057 405 LIB3059-048- LIB3059 g22485 BLASTN 1822 1e−142 99 Q1-K1-H5 1058 405 LIB3078-040- LIB3078 g514945 BLASTN 1801 1e−141 97 Q1-K1-F8 1059 405 LIB3078-001- LIB3078 g22485 BLASTN 1246 1e−139 95 Q1-K1-C7 1060 405 LIB3061-024- LIB3061 g22485 BLASTN 1376 1e−139 94 Q1-K1-A12 1061 405 LIB3061-026- LIB3061 g22485 BLASTN 1643 1e−138 93 Q1-K1-D3 1062 405 LIB3061-056- LIB3061 g22485 BLASTN 1763 1e−138 92 Q1-K1-D8 1063 405 LIB3069-041- LIB3069 g514945 BLASTN 1758 1e−137 97 Q1-K1-G12 1064 405 LIB3059-025- LIB3059 g22485 BLASTN 1532 1e−132 94 Q1-K1-E5 1065 405 LIB3061-014- LIB3061 g22485 BLASTN 1294 1e−130 88 Q1-K1-D4 1066 405 LIB3061-005- LIB3061 g22485 BLASTN 1540 1e−130 97 Q1-K1-C9 1067 405 LIB3061-016- LIB3061 g22485 BLASTN 1251 1e−129 85 Q1-K1-G2 1068 405 LIB3069-029- LIB3069 g514945 BLASTN 1657 1e−129 88 Q1-K1-B2 1069 405 LIB3078-012- LIB3078 g514945 BLASTN 857 1e−128 86 Q1-K1-F7 1070 405 LIB3078-016- LIB3078 g514945 BLASTN 1335 1e−128 87 Q1-K1-D7 1071 405 LIB3062-049- LIB3062 g514945 BLASTN 1609 1e−128 88 Q1-K1-A8 1072 405 LIB143-006- LIB143 g514945 BLASTN 1614 1e−125 96 Q1-E1-G12 1073 405 LIB3059-024- LIB3059 g22485 BLASTN 1529 1e−123 83 Q1-K1-E5 1074 405 LIB3069-008- LIB3069 g514945 BLASTN 1036 1e−115 94 Q1-K1-C1 1075 405 LIB3059-018- LIB3059 g514945 BLASTN 910 1e−103 93 Q1-K1-F11 1076 405 LIB3078-001- LIB3078 g514945 BLASTN 952 1e−98 90 Q1-K1-E8 1077 405 LIB3059-017- LIB3059 g22485 BLASTN 1170 1e−88 92 Q1-K1-G4 1078 405 LIB3067-045- LIB3067 g533251 BLASTN 917 1e−87 87 Q1-K1-E9 1079 405 LIB3062-015- LIB3062 g514945 BLASTN 1066 1e−86 96 Q1-K1-C1 1080 405 LIB3059-039- LIB3059 g22485 BLASTN 856 1e−82 92 Q1-K1-A3 1081 405 LIB3062-024- LIB3062 g514945 BLASTN 548 1e−79 88 Q1-K1-C3 1082 405 LIB3059-029- LIB3059 g22485 BLASTN 925 1e−74 94 Q1-K1-F1 1083 405 LIB3059-006- LIB3059 g22485 BLASTN 530 1e−50 83 Q1-K1-F4 1084 405 LIB3067-017- LIB3067 g533251 BLASTN 425 1e−26 100 Q1-K1-C3 1085 405 LIB3061-028- LIB3061 g19106 BLASTX 118 1e−25 100 Q1-K1-A9 1086 537 LIB3066-009- LIB3066 g22485 BLASTN 1369 1e−122 96 Q1-K1-B9 MAIZE HEXOKINASE 1087 −700018381 700018381H1 SATMON001 g1899025 BLASTX 166 1e−16 48 1088 −700051079 700051079H1 SATMON003 g1899025 BLASTX 84 1e−11 50 1089 −700101579 700101579H1 SATMON009 g881521 BLASTX 217 1e−23 66 1090 −700105594 700105594H1 SATMON010 g3087888 BLASTX 181 1e−17 57 1091 −700106018 700106018H1 SATMON010 g3087888 BLASTX 195 1e−19 64 1092 −700157233 700157233H1 SATMON012 g3087888 BLASTX 198 1e−20 58 1093 −700202992 700202992H1 SATMON003 g3087888 BLASTX 89 1e−9 58 1094 −700224204 700224204H1 SATMON011 g1899024 BLASTN 520 1e−34 70 1095 −700241273 700241273H1 SATMON010 g3087888 BLASTX 184 1e−18 58 1096 −700352183 700352183H1 SATMON023 g1899024 BLASTN 481 1e−31 70 1097 −700573814 700573814H1 SATMON030 g1899024 BLASTN 535 1e−34 67 1098 −700612458 700612458H1 SATMON033 g619928 BLASTX 229 1e−26 61 1099 −701168774 701168774H1 SATMONN05 g619927 BLASTN 252 1e−10 62 1100 1195 700457430H1 SATMON029 g3087888 BLASTX 122 1e−19 53 1101 13262 700102942H1 SATMON010 g3087888 BLASTX 113 1e−18 53 1102 1378 700456148H1 SATMON029 g1899025 BLASTX 267 1e−29 59 1103 1378 700455837H1 SATMON029 g1899025 BLASTX 166 1e−21 60 1104 17305 700460742H1 SATMON031 g619928 BLASTX 131 1e−15 57 1105 17305 700614972H1 SATMON033 g1899025 BLASTX 100 1e−8 53 1106 1842 700089135H1 SATMON011 g619928 BLASTX 405 1e−49 70 1107 1842 700430234H1 SATMONN01 g619927 BLASTN 461 1e−28 72 1108 1842 700166122H1 SATMON013 g619928 BLASTX 183 1e−18 84 1109 24376 700053677H1 SATMON010 g1899024 BLASTN 642 1e−44 70 1110 24376 700152328H1 SATMON007 g619927 BLASTN 555 1e−37 69 1111 24376 700623451H1 SATMON034 g619928 BLASTX 197 1e−32 72 1112 28388 700089065H1 SATMON011 g619928 BLASTX 186 1e−30 61 1113 3345 700072110H1 SATMON007 g619928 BLASTX 125 1e−24 66 1114 3345 700472061H1 SATMON025 g619928 BLASTX 112 1e−20 55 1115 3345 701173753H1 SATMONN05 g619928 BLASTX 135 1e−16 54 1116 3345 700202130H1 SATMON003 g619928 BLASTX 113 1e−11 68 1117 5073 700582054H1 SATMON031 g619928 BLASTX 247 1e−29 66 1118 5073 700053432H1 SATMON009 g619928 BLASTX 233 1e−25 60 1119 6731 700099009H1 SATMON009 g619927 BLASTN 736 1e−52 72 1120 6731 700089738H1 SATMON011 g1899024 BLASTN 700 1e−49 70 1121 6731 700171542H1 SATMON013 g619927 BLASTN 530 1e−35 74 1122 7565 700356773H1 SATMON024 g1899025 BLASTX 177 1e−17 62 1123 9695 700212172H1 SATMON016 g1899024 BLASTN 832 1e−60 74 1124 9695 700212124H1 SATMON016 g1899024 BLASTN 835 1e−60 75 1125 9695 700094278H1 SATMON008 g1899024 BLASTN 819 1e−59 74 1126 −L30621307 LIB3062-001- LIB3062 g1899025 BLASTX 95 1e−32 53 Q1-K2-G11 1127 −L30782665 LIB3078-007- LIB3078 g3087888 BLASTX 130 1e−39 47 Q1-K1-E9 1128 24376 LIB3069-041- LIB3069 g1899024 BLASTN 608 1e−61 70 Q1-K1-E7 1129 28244 LIB3061-004- LIB3061 g687676 BLASTN 499 1e−30 65 Q1-K1-F9 1130 28388 LIB3066-030- LIB3066 g619928 BLASTX 299 1e−63 64 Q1-K1-G10 1131 3364 LIB3078-051- LIB3078 g687676 BLASTN 619 1e−41 67 Q1-K1-B3 1132 3364 LIB3078-053- LIB3078 g687676 BLASTN 627 1e−41 69 Q1-K1-C9 1133 3364 LIB84-015- LIB84 g687676 BLASTN 554 1e−35 69 Q1-E1-F7 1134 6731 LIB3061-028- LIB3061 g1899024 BLASTN 831 1e−60 70 Q1-K1-C1 1135 9695 LIB143-065- LIB143 g1899024 BLASTN 1096 1e−82 73 Q1-E1-C10 MAIZE FRUCTOKINASE 1136 −700106058 700106058H1 SATMON010 g1052972 BLASTN 220 1e−9 68 1137 −700151135 700151135H1 SATMON007 g297014 BLASTN 351 1e−18 75 1138 −700169310 700169310H1 SATMON013 g1052972 BLASTN 273 1e−12 59 1139 −700210226 700210226H1 SATMON016 g1052973 BLASTX 188 1e−24 68 1140 −700257901 700257901H1 SATMON017 g297015 BLASTX 200 1e−20 72 1141 −700621274 700621274H1 SATMON034 g1052973 BLASTX 141 1e−24 64 1142 11678 700105513H1 SATMON010 g1052972 BLASTN 580 1e−39 64 1143 11678 700170725H1 SATMON013 g1052972 BLASTN 478 1e−31 66 1144 2526 700159958H1 SATMON012 g1052973 BLASTX 152 1e−14 64 1145 2754 700102678H1 SATMON010 g1052972 BLASTN 707 1e−50 69 1146 2754 700102312H1 SATMON010 g1052972 BLASTN 701 1e−49 69 1147 2754 700205695H1 SATMON003 g1915973 BLASTN 633 1e−43 69 1148 2754 700221511H1 SATMON011 g1915973 BLASTN 587 1e−40 69 1149 2754 700469079H1 SATMON025 g1052972 BLASTN 584 1e−39 72 1150 2754 701173520H1 SATMONN05 g1915973 BLASTN 342 1e−36 70 1151 2754 700267332H1 SATMON017 g1052972 BLASTN 541 1e−35 64 1152 2754 701164907H1 SATMONN04 g1052973 BLASTX 280 1e−33 57 1153 2754 700450050H2 SATMON028 g1052973 BLASTX 160 1e−31 60 1154 2754 701182860H1 SATMONN06 g297015 BLASTX 188 1e−27 65 1155 2754 700467520H1 SATMON025 g1915974 BLASTX 242 1e−26 60 1156 2754 700159848H1 SATMON012 g1052973 BLASTX 197 1e−24 63 1157 3287 700088103H1 SATMON011 g2102693 BLASTX 239 1e−43 74 1158 3287 700210913H1 SATMON016 g2102693 BLASTX 250 1e−35 77 1159 3287 700167609H1 SATMON013 g1052973 BLASTX 300 1e−35 68 1160 3287 700085916H1 SATMON011 g1052972 BLASTN 553 1e−35 64 1161 3287 700262715H1 SATMON017 g1915974 BLASTX 201 1e−33 71 1162 3287 700170179H1 SATMON013 g1052973 BLASTX 289 1e−33 67 1163 3287 700615671H1 SATMON033 g1052972 BLASTN 515 1e−32 63 1164 3287 700223640H1 SATMON011 g1052973 BLASTX 219 1e−31 67 1165 3287 700215234H1 SATMON016 g1052973 BLASTX 190 1e−30 67 1166 3287 700203946H1 SATMON003 g1052973 BLASTX 198 1e−30 60 1167 3287 700028411H1 SATMON003 g2102693 BLASTX 110 1e−29 57 1168 3287 700224307H1 SATMON011 g1052973 BLASTX 159 1e−29 87 1169 3287 700072013H1 SATMON007 g1052973 BLASTX 191 1e−29 65 1170 3287 700215669H1 SATMON016 g1052973 BLASTX 260 1e−29 57 1171 3287 700353954H1 SATMON024 g1052973 BLASTX 260 1e−29 61 1172 3287 700342211H1 SATMON021 g1052973 BLASTX 137 1e−28 67 1173 3287 700085462H1 SATMON011 g297014 BLASTN 466 1e−28 62 1174 3287 700220972H1 SATMON011 g1052973 BLASTX 109 1e−27 83 1175 3287 700451141H1 SATMON028 g1052973 BLASTX 245 1e−27 63 1176 3287 700087484H1 SATMON011 g1052972 BLASTN 440 1e−26 64 1177 3287 700343411H1 SATMON021 g1052973 BLASTX 163 1e−25 67 1178 3287 700217263H1 SATMON016 g1915973 BLASTN 393 1e−25 68 1179 3287 700030665H1 SATMON003 g1052973 BLASTX 176 1e−24 71 1180 3287 700343380H1 SATMON021 g1052973 BLASTX 228 1e−24 57 1181 3287 701159743H2 SATMONN04 g1052973 BLASTX 183 1e−23 55 1182 3287 700221543H1 SATMON011 g1052973 BLASTX 217 1e−23 50 1183 3287 700333946H1 SATMON019 g1052973 BLASTX 178 1e−22 66 1184 3287 700091730H1 SATMON011 g1052973 BLASTX 171 1e−21 64 1185 3287 700570521H1 SATMON030 g1915974 BLASTX 98 1e−18 58 1186 3287 700048604H1 SATMON003 g1052973 BLASTX 88 1e−15 54 1187 3287 700208681H1 SATMON016 g1052973 BLASTX 129 1e−15 55 1188 3287 700028328H1 SATMON003 g1052973 BLASTX 162 1e−15 66 1189 3287 700220530H1 SATMON011 g1052973 BLASTX 141 1e−14 88 1190 3287 700243726H1 SATMON010 g1052973 BLASTX 153 1e−14 68 1191 3287 700142502H1 SATMON012 g1052973 BLASTX 157 1e−14 47 1192 3287 700336537H1 SATMON019 g1052973 BLASTX 141 1e−12 50 1193 3287 700205308H1 SATMON003 g1052973 BLASTX 133 1e−11 75 1194 5966 700084171H1 SATMON011 g1052972 BLASTN 448 1e−26 66 1195 5966 700084951H1 SATMON011 g2102693 BLASTX 214 1e−22 73 1196 5966 700089353H1 SATMON011 g2102691 BLASTX 195 1e−20 72 1197 5966 700220723H1 SATMON011 g1915974 BLASTX 198 1e−20 73 1198 5966 700084412H1 SATMON011 g2102693 BLASTX 179 1e−19 76 1199 5966 700085628H1 SATMON011 g2102691 BLASTX 180 1e−18 72 1200 5966 700027982H1 SATMON003 g2102691 BLASTX 178 1e−17 72 1201 5966 700106884H1 SATMON010 g1915974 BLASTX 148 1e−13 75 1202 5966 700053135H1 SATMON008 g1915974 BLASTX 131 1e−11 73 1203 5966 700027988H1 SATMON003 g1915974 BLASTX 134 1e−11 65 1204 5966 700207083H1 SATMON003 g1915974 BLASTX 100 1e−10 46 1205 5966 700158574H1 SATMON012 g1915974 BLASTX 120 1e−9 50 1206 2754 LIB3061-030- LIB3061 g1052972 BLASTN 882 1e−64 67 Q1-K1-G12 1207 2754 LIB3061-030- LIB3061 g1052972 BLASTN 751 1e−52 68 Q1-K1-G11 1208 3287 LIB3067-040- LIB3067 g1052972 BLASTN 657 1e−44 64 Q1-K1-H10 1209 3287 LIB84-024- LIB84 g1052972 BLASTN 638 1e−42 64 Q1-E1-H7 1210 3287 LIB3069-045- LIB3069 g1052972 BLASTN 592 1e−38 61 Q1-K1-F6 1211 3287 LIB3061-014- LIB3061 g1052973 BLASTX 175 1e−36 41 Q1-K1-A3 1212 3287 LIB3062-019- LIB3062 g1052973 BLASTX 154 1e−30 68 Q1-K1-H11 1213 3287 LIB3067-054- LIB3067 g1052972 BLASTN 495 1e−30 61 Q1-K1-C9 1214 3287 LIB3067-022- LIB3067 g1052973 BLASTX 141 1e−27 68 Q1-K1-H4 1215 3287 LIB3069-045- LIB3069 g1052972 BLASTN 439 1e−25 57 Q1-K1-F2 MAIZE NDP-KINASE 1216 −700575072 700575072H1 SATMON030 g303849 BLASTX 74 1e−13 89 1217 −701170773 701170773H1 SATMONN05 g1777930 BLASTX 132 1e−30 71 1218 2462 700050003H1 SATMON003 g218233 BLASTN 656 1e−58 83 1219 2462 700204789H1 SATMON003 g218233 BLASTN 780 1e−58 87 1220 2462 700049819H1 SATMON003 g218233 BLASTN 786 1e−58 86 1221 2462 700204211H1 SATMON003 g218233 BLASTN 786 1e−58 86 1222 2462 700205742H1 SATMON003 g218233 BLASTN 763 1e−57 86 1223 2462 700207611H1 SATMON016 g218233 BLASTN 764 1e−57 87 1224 2462 700072505H1 SATMON007 g218233 BLASTN 740 1e−55 86 1225 2462 700236468H1 SATMON010 g218233 BLASTN 710 1e−52 86 1226 2462 701181270H1 SATMONN06 g218233 BLASTN 445 1e−51 86 1227 2462 700573201H1 SATMON030 g218233 BLASTN 691 1e−51 81 1228 2462 700452623H1 SATMON028 g218233 BLASTN 694 1e−51 85 1229 2462 700351523H1 SATMON023 g218233 BLASTN 679 1e−50 86 1230 2462 700042795H1 SATMON004 g218233 BLASTN 630 1e−45 86 1231 2462 700445979H1 SATMON027 g218233 BLASTN 595 1e−43 86 1232 2462 700201855H1 SATMON003 g218233 BLASTN 604 1e−43 87 1233 2462 700573101H1 SATMON030 g218233 BLASTN 594 1e−42 78 1234 2462 700049543H1 SATMON003 g218233 BLASTN 577 1e−41 79 1235 2462 700432359H1 SATMONN01 g218233 BLASTN 561 1e−40 81 1236 2462 701182021H1 SATMONN06 g218233 BLASTN 561 1e−40 85 1237 2462 701182019H1 SATMONN06 g218233 BLASTN 566 1e−40 86 1238 2462 700150928H1 SATMON007 g218233 BLASTN 569 1e−40 85 1239 2462 700202824H1 SATMON003 g218233 BLASTN 336 1e−39 86 1240 2462 700451056H1 SATMON028 g218233 BLASTN 553 1e−39 85 1241 2462 700449958H1 SATMON028 g218233 BLASTN 544 1e−38 86 1242 2462 700347592H1 SATMON023 g218233 BLASTN 403 1e−34 78 1243 2462 700573195H1 SATMON030 g218233 BLASTN 200 1e−22 84 1244 2462 700582836H1 SATMON031 g303849 BLASTX 157 1e−15 83 1245 2462 700029459H1 SATMON003 g303849 BLASTX 134 1e−11 84 1246 27065 700583429H1 SATMON031 g1064895 BLASTX 72 1e−13 54 1247 −L1482546 LIB148-007- LIB148 g218233 BLASTN 359 1e−19 75 Q1-E1-E6 1248 2462 LIB3067-039- LIB3067 g218233 BLASTN 711 1e−52 82 Q1-K1-B10 1249 2462 LIB3078-001- LIB3078 g218233 BLASTN 488 1e−49 85 Q1-K1-F3 1250 2462 LIB3067-029- LIB3067 g1236951 BLASTX 166 1e−31 96 Q1-K1-C3 1251 25174 LIB189-022- LIB189 g758643 BLASTN 440 1e−25 76 Q1-E1-E9 MAIZE GLUCOSE-6-PHOSPHATE 1-DEHYDROGENASE 1252 −700047645 700047645H1 SATMON003 g471345 BLASTX 193 1e−21 58 1253 −700210379 700210379H1 SATMON016 g1480344 BLASTX 103 1e−10 85 1254 9135 700203121H1 SATMON003 g1166405 BLASTX 108 1e−10 78 MAIZE PHOSPHOGLUCOMUTASE 1255 −700045655 700045655H1 SATMON004 g534982 BLASTX 144 1e−12 65 1256 −700053330 700053330H1 SATMON009 g3294467 BLASTX 211 1e−23 71 1257 −700102193 700102193H1 SATMON010 g534982 BLASTX 145 1e−14 53 1258 −700166982 700166982H1 SATMON013 g2795876 BLASTX 168 1e−16 52 1259 −700169540 700169540H1 SATMON013 g534982 BLASTX 180 1e−17 61 1260 −700210088 700210088H1 SATMON016 g534982 BLASTX 328 1e−38 55 1261 −700573194 700573194H1 SATMON030 g534982 BLASTX 192 1e−21 54 1262 −700616588 700616588H1 SATMON033 g3294468 BLASTN 593 1e−48 95 1263 119 700574655H1 SATMON030 g3294466 BLASTN 1705 1e−133 98 1264 119 700574672H1 SATMON030 g3294466 BLASTN 820 1e−121 100 1265 119 700100992H1 SATMON009 g3294466 BLASTN 1545 1e−119 99 1266 119 700615409H1 SATMON033 g3294466 BLASTN 1050 1e−118 100 1267 119 700210693H1 SATMON016 g3294468 BLASTN 1515 1e−117 100 1268 119 700381526H1 SATMON023 g3294468 BLASTN 1490 1e−115 100 1269 119 700026372H1 SATMON003 g3294466 BLASTN 1463 1e−113 99 1270 119 700201578H1 SATMON003 g3294468 BLASTN 677 1e−112 96 1271 119 700101083H1 SATMON009 g3294468 BLASTN 1430 1e−110 100 1272 119 700217101H1 SATMON016 g3294468 BLASTN 1420 1e−109 100 1273 119 700222466H1 SATMON011 g3294466 BLASTN 957 1e−106 97 1274 119 700072492H1 SATMON007 g3294466 BLASTN 1381 1e−106 99 1275 119 700043724H1 SATMON004 g3294468 BLASTN 1390 1e−106 100 1276 119 700346762H1 SATMON021 g3294468 BLASTN 1333 1e−102 94 1277 119 700347741H1 SATMON023 g3294468 BLASTN 1339 1e−102 97 1278 119 700550792H1 SATMON022 g3294466 BLASTN 731 1e−101 99 1279 119 700380144H1 SATMON021 g3294466 BLASTN 1216 1e−98 97 1280 119 700241526H1 SATMON010 g3294466 BLASTN 1285 1e−98 100 1281 119 700380456H1 SATMON021 g3294468 BLASTN 650 1e−97 99 1282 119 700238734H1 SATMON010 g3294466 BLASTN 974 1e−97 97 1283 119 700083634H1 SATMON011 g3294468 BLASTN 1265 1e−96 100 1284 119 700383086H1 SATMON024 g3294466 BLASTN 961 1e−94 96 1285 119 700169630H1 SATMON013 g3294466 BLASTN 1245 1e−94 100 1286 119 701177766H1 SATMONN05 g3294466 BLASTN 1187 1e−93 97 1287 119 700142461H1 SATMON012 g3294466 BLASTN 1231 1e−93 99 1288 119 700044235H1 SATMON004 g3294466 BLASTN 1175 1e−89 100 1289 119 700216921H1 SATMON016 g3294466 BLASTN 1165 1e−88 100 1290 119 700333779H1 SATMON019 g3294466 BLASTN 996 1e−87 96 1291 119 700021881H1 SATMON001 g3294468 BLASTN 1120 1e−84 100 1292 119 700049194H1 SATMON003 g3294468 BLASTN 940 1e−82 98 1293 119 700164477H1 SATMON013 g3294466 BLASTN 1091 1e−82 99 1294 119 700169514H1 SATMON013 g3294468 BLASTN 865 1e−80 100 1295 119 700050896H1 SATMON003 g3294466 BLASTN 591 1e−76 94 1296 119 700172394H1 SATMON013 g3294466 BLASTN 1024 1e−76 99 1297 119 700211437H1 SATMON016 g3294466 BLASTN 994 1e−73 99 1298 119 700084535H1 SATMON011 g3294468 BLASTN 973 1e−72 99 1299 119 700203439H1 SATMON003 g3294466 BLASTN 765 1e−71 100 1300 119 700257833H1 SATMON017 g3294468 BLASTN 611 1e−69 94 1301 119 700621831H1 SATMON034 g3294466 BLASTN 412 1e−52 90 1302 119 700354511H1 SATMON024 g3294468 BLASTN 703 1e−52 91 1303 119 700203525H1 SATMON003 g3294468 BLASTN 708 1e−50 99 1304 119 700020476H1 SATMON001 g3294468 BLASTN 658 1e−45 99 1305 119 700050562H1 SATMON003 g3294466 BLASTN 544 1e−42 88 1306 119 700613868H1 SATMON033 g3294466 BLASTN 615 1e−42 100 1307 119 700574982H1 SATMON030 g3294466 BLASTN 473 1e−35 97 1308 119 700049512H1 SATMON003 g3294466 BLASTN 268 1e−29 95 1309 119 700260372H2 SATMON017 g3294466 BLASTN 226 1e−10 89 1310 16726 700082801H1 SATMON011 g2829893 BLASTX 278 1e−30 55 1311 16726 700212054H1 SATMON016 g2829893 BLASTX 220 1e−23 53 1312 19462 700097450H1 SATMON009 g1814400 BLASTN 323 1e−29 64 1313 19462 700441165H1 SATMON026 g1408296 BLASTX 239 1e−25 61 1314 24348 700379424H1 SATMON020 g3294466 BLASTN 707 1e−50 98 1315 2587 700089556H1 SATMON011 g2829893 BLASTX 117 1e−8 67 1316 3016 700204345H1 SATMON003 g3294468 BLASTN 1784 1e−139 98 1317 3016 700098713H1 SATMON009 g3294468 BLASTN 1516 1e−117 99 1318 3016 700084751H1 SATMON011 g3294466 BLASTN 1475 1e−114 100 1319 3016 700351326H1 SATMON023 g3294468 BLASTN 1460 1e−112 100 1320 3016 700097161H1 SATMON009 g3294466 BLASTN 1308 1e−109 98 1321 3016 700266423H1 SATMON017 g3294468 BLASTN 1065 1e−108 96 1322 3016 700349605H1 SATMON023 g3294466 BLASTN 1335 1e−107 100 1323 3016 700350209H1 SATMON023 g3294468 BLASTN 1188 1e−106 97 1324 3016 700265291H1 SATMON017 g3294468 BLASTN 873 1e−100 98 1325 3016 700457572H1 SATMON029 g3294466 BLASTN 1288 1e−98 98 1326 3016 700334810H1 SATMON019 g3294468 BLASTN 863 1e−97 99 1327 3016 700194444H1 SATMON014 g3294466 BLASTN 1265 1e−96 100 1328 3016 700457426H1 SATMON029 g3294466 BLASTN 1236 1e−94 98 1329 3016 700210958H1 SATMON016 g3294466 BLASTN 1148 1e−92 98 1330 3016 700075135H1 SATMON007 g3294468 BLASTN 1219 1e−92 97 1331 3016 700152065H1 SATMON007 g3294466 BLASTN 1135 1e−90 99 1332 3016 700219672H1 SATMON011 g3294468 BLASTN 823 1e−89 99 1333 3016 700170425H1 SATMON013 g3294466 BLASTN 1110 1e−83 100 1334 3016 700153495H1 SATMON007 g3294468 BLASTN 640 1e−82 100 1335 3016 700348567H1 SATMON023 g3294468 BLASTN 557 1e−81 87 1336 3016 700803158H1 SATMON036 g3294468 BLASTN 630 1e−60 85 1337 3016 700264923H1 SATMON017 g3294468 BLASTN 340 1e−50 98 1338 3016 700615715H1 SATMON033 g3294466 BLASTN 567 1e−48 96 1339 3016 700027830H1 SATMON003 g3294468 BLASTN 632 1e−43 95 1340 3016 700350539H1 SATMON023 g3294466 BLASTN 333 1e−41 96 1341 4562 700044891H1 SATMON004 g3294466 BLASTN 650 1e−45 74 1342 4562 700215538H1 SATMON016 g3294466 BLASTN 555 1e−37 67 1343 9894 700220429H1 SATMON011 g3294468 BLASTN 1302 1e−99 99 1344 9894 700236461H1 SATMON010 g3294466 BLASTN 1054 1e−90 97 1345 −L30594453 LIB3059-042- LIB3059 g1814401 BLASTX 290 1e−49 58 Q1-K1-B5 1346 −L30605287 LIB3060-049- LIB3060 g534982 BLASTX 172 1e−34 77 Q1-K1-B7 1347 119 LIB3059-019- LIB3059 g1881692 BLASTN 2094 1e−165 98 Q1-K1-H1 1348 119 LIB3059-031- LIB3059 g1881692 BLASTN 1926 1e−151 96 Q1-K1-H10 1349 119 LIB3069-012- LIB3069 g1881692 BLASTN 1188 1e−146 90 Q1-K1-F2 1350 119 LIB36-019- LIB36 g1881692 BLASTN 1783 1e−139 90 Q1-E1-A7 1351 119 LIB3078-023- LIB3078 g1881692 BLASTN 860 1e−124 87 Q1-K1-C3 1352 119 LIB3067-058- LIB3067 g1881692 BLASTN 991 1e−114 99 Q1-K1-G1 1353 119 LIB3062-048- LIB3062 g1881692 BLASTN 1181 1e−103 97 Q1-K1-B7 1354 119 LIB3069-023- LIB3069 g1881692 BLASTN 1176 1e−87 84 Q1-K1-G4 1355 119 LIB3069-025- LIB3069 g1881692 BLASTN 611 1e−65 91 Q1-K1-B6 1356 24348 LIB3066-043- LIB3066 g1881692 BLASTN 560 1e−37 100 Q1-K1-F11 1357 24348 LIB3067-048- LIB3067 g1881692 BLASTN 543 1e−36 99 Q1-K1-F3 1358 3016 LIB143-002- LIB143 g2829893 BLASTX 224 1e−51 72 Q1-E1-C12 1359 3016 LIB189-034- LIB189 g2829893 BLASTX 216 1e−48 68 Q1-E1-A11 1360 3016 LIB3069-043- LIB3069 g1814401 BLASTX 98 1e−32 64 Q1-K1-D5 MAIZE UDP-GLUCOSE PYROPHOSPHORYLASE 1361 −700197315 700197315H1 SATMON014 g1388021 BLASTX 122 1e−9 70 1362 −700203530 700203530H1 SATMON003 g1212995 BLASTN 568 1e−38 78 1363 −700267284 700267284H1 SATMON017 g1212996 BLASTX 150 1e−13 87 1364 −700336683 700336683H1 SATMON019 g1752677 BLASTX 150 1e−27 82 1365 −700342324 700342324H1 SATMON021 g3107931 BLASTX 95 1e−14 80 1366 −700354856 700354856H1 SATMON024 g1388021 BLASTX 121 1e−22 75 1367 −700613858 700613858H1 SATMON033 g1212995 BLASTN 776 1e−59 88 1368 14982 700028996H1 SATMON003 g1212995 BLASTN 560 1e−37 76 1369 14982 700155115H1 SATMON007 g1212995 BLASTN 399 1e−31 81 1370 14982 700356747H1 SATMON024 g1388021 BLASTX 166 1e−15 76 1371 19537 700573761H1 SATMON030 g1212995 BLASTN 954 1e−70 79 1372 19537 700208049H1 SATMON016 g1212995 BLASTN 901 1e−66 78 1373 19537 700086382H1 SATMON011 g1212995 BLASTN 885 1e−64 77 1374 69 700091881H1 SATMON011 g1212995 BLASTN 844 1e−105 89 1375 69 700624406H1 SATMON034 g1212995 BLASTN 816 1e−97 88 1376 69 700211464H1 SATMON016 g1212995 BLASTN 1251 1e−95 88 1377 69 700099836H1 SATMON009 g1212995 BLASTN 1239 1e−94 88 1378 69 700084756H1 SATMON011 g1212995 BLASTN 1240 1e−94 90 1379 69 700076136H1 SATMON007 g1212995 BLASTN 1243 1e−94 89 1380 69 700073071H1 SATMON007 g1212995 BLASTN 1163 1e−88 86 1381 69 700614228H1 SATMON033 g1212995 BLASTN 1013 1e−87 84 1382 69 700379926H1 SATMON021 g1212995 BLASTN 1138 1e−86 88 1383 69 700089172H1 SATMON011 g1212995 BLASTN 1141 1e−86 88 1384 69 700265063H1 SATMON017 g1212995 BLASTN 1147 1e−86 86 1385 69 700085964H1 SATMON011 g1212995 BLASTN 1135 1e−85 85 1386 69 700282281H2 SATMON023 g1212995 BLASTN 1136 1e−85 86 1387 69 700429855H1 SATMONN01 g1212995 BLASTN 1114 1e−84 89 1388 69 700347453H1 SATMON023 g1212995 BLASTN 1117 1e−84 87 1389 69 700265087H1 SATMON017 g1212995 BLASTN 1120 1e−84 87 1390 69 700092705H1 SATMON008 g1212995 BLASTN 1122 1e−84 87 1391 69 700212686H1 SATMON016 g1212995 BLASTN 1123 1e−84 91 1392 69 700623332H1 SATMON034 g1212995 BLASTN 800 1e−83 86 1393 69 700041787H1 SATMON004 g1212995 BLASTN 1091 1e−82 91 1394 69 700219031H1 SATMON011 g1212995 BLASTN 1093 1e−82 89 1395 69 700218632H1 SATMON011 g1212995 BLASTN 1086 1e−81 90 1396 69 700211962H1 SATMON016 g1212995 BLASTN 1086 1e−81 86 1397 69 700220729H1 SATMON011 g1212995 BLASTN 916 1e−80 84 1398 69 700197025H1 SATMON014 g1212995 BLASTN 1063 1e−79 89 1399 69 700086546H1 SATMON011 g1212995 BLASTN 1049 1e−78 86 1400 69 700217064H1 SATMON016 g1212995 BLASTN 1051 1e−78 88 1401 69 700799128H1 SATMON036 g1212995 BLASTN 618 1e−77 88 1402 69 700265488H1 SATMON017 g1212995 BLASTN 1030 1e−77 84 1403 69 700043842H1 SATMON004 g1212995 BLASTN 1035 1e−77 87 1404 69 700236833H1 SATMON010 g1212995 BLASTN 1035 1e−77 87 1405 69 700219083H1 SATMON011 g1212995 BLASTN 1036 1e−77 88 1406 69 700042338H1 SATMON004 g1212995 BLASTN 1037 1e−77 87 1407 69 700352484H1 SATMON023 g1212995 BLASTN 1038 1e−77 85 1408 69 700083771H1 SATMON011 g1212995 BLASTN 613 1e−76 91 1409 69 700473855H1 SATMON025 g1212995 BLASTN 755 1e−76 85 1410 69 700353922H1 SATMON024 g1212995 BLASTN 1024 1e−76 85 1411 69 700023267H1 SATMON003 g1212995 BLASTN 1007 1e−75 89 1412 69 700157596H1 SATMON012 g1212995 BLASTN 1008 1e−75 87 1413 69 700218718H1 SATMON011 g1212995 BLASTN 1012 1e−75 86 1414 69 700162316H1 SATMON012 g1212995 BLASTN 626 1e−74 80 1415 69 700046475H1 SATMON004 g1212995 BLASTN 1003 1e−74 85 1416 69 700466010H1 SATMON025 g1212995 BLASTN 558 1e−73 82 1417 69 700571392H1 SATMON030 g1212995 BLASTN 985 1e−73 85 1418 69 700165241H1 SATMON013 g1212995 BLASTN 987 1e−73 85 1419 69 700457410H1 SATMON029 g1212995 BLASTN 988 1e−73 87 1420 69 700194672H1 SATMON014 g1212995 BLASTN 963 1e−71 86 1421 69 700089746H1 SATMON011 g1212995 BLASTN 964 1e−71 83 1422 69 700801620H1 SATMON036 g1212995 BLASTN 536 1e−70 91 1423 69 700264785H1 SATMON017 g1212995 BLASTN 952 1e−70 84 1424 69 700244093H1 SATMON010 g1212995 BLASTN 954 1e−70 85 1425 69 700043787H1 SATMON004 g1212995 BLASTN 957 1e−70 85 1426 69 700267269H1 SATMON017 g1212995 BLASTN 867 1e−69 85 1427 69 700167985H1 SATMON013 g1212995 BLASTN 940 1e−69 89 1428 69 700799042H1 SATMON036 g1212995 BLASTN 812 1e−68 89 1429 69 700163824H1 SATMON013 g1212995 BLASTN 888 1e−65 86 1430 69 700098307H1 SATMON009 g1212995 BLASTN 461 1e−63 81 1431 69 700805267H1 SATMON036 g1212995 BLASTN 734 1e−63 88 1432 69 700204843H1 SATMON003 g1212995 BLASTN 854 1e−62 88 1433 69 700206721H1 SATMON003 g1212995 BLASTN 461 1e−61 81 1434 69 700018559H1 SATMON001 g1212995 BLASTN 847 1e−61 85 1435 69 700026241H1 SATMON003 g1212995 BLASTN 847 1e−61 87 1436 69 700099987H1 SATMON009 g1212995 BLASTN 461 1e−60 81 1437 69 700475628H1 SATMON025 g1212995 BLASTN 750 1e−59 80 1438 69 700016675H1 SATMON001 g1212995 BLASTN 814 1e−59 86 1439 69 700150144H1 SATMON007 g1212995 BLASTN 822 1e−59 86 1440 69 700267260H1 SATMON017 g1212995 BLASTN 461 1e−58 80 1441 69 700261336H1 SATMON017 g1212995 BLASTN 564 1e−58 82 1442 69 700618652H1 SATMON033 g1212995 BLASTN 730 1e−58 78 1443 69 700469914H1 SATMON025 g1212995 BLASTN 735 1e−58 89 1444 69 700048027H1 SATMON003 g1212995 BLASTN 807 1e−58 83 1445 69 700165703H1 SATMON013 g1212995 BLASTN 796 1e−57 85 1446 69 700265403H1 SATMON017 g1212995 BLASTN 797 1e−57 78 1447 69 700099428H1 SATMON009 g1212995 BLASTN 474 1e−56 88 1448 69 700243212H1 SATMON010 g1212995 BLASTN 779 1e−56 84 1449 69 700092996H1 SATMON008 g1212995 BLASTN 789 1e−56 84 1450 69 700803035H1 SATMON036 g1212995 BLASTN 436 1e−54 80 1451 69 700235803H1 SATMON010 g1212995 BLASTN 688 1e−54 79 1452 69 700172581H1 SATMON013 g1212995 BLASTN 754 1e−54 79 1453 69 700214715H1 SATMON016 g1212995 BLASTN 762 1e−54 86 1454 69 700223082H1 SATMON011 g1212995 BLASTN 764 1e−54 84 1455 69 700093483H1 SATMON008 g1212995 BLASTN 357 1e−51 88 1456 69 700261920H1 SATMON017 g1212995 BLASTN 363 1e−51 82 1457 69 700221718H1 SATMON011 g1212995 BLASTN 363 1e−51 83 1458 69 700453106H1 SATMON028 g1212995 BLASTN 670 1e−51 82 1459 69 700210506H1 SATMON016 g1212995 BLASTN 461 1e−50 85 1460 69 700212333H1 SATMON016 g1212995 BLASTN 443 1e−49 83 1461 69 700072654H1 SATMON007 g1212995 BLASTN 443 1e−49 79 1462 69 700218282H1 SATMON016 g1212995 BLASTN 452 1e−49 85 1463 69 700263725H1 SATMON017 g1212995 BLASTN 662 1e−49 80 1464 69 700343083H1 SATMON021 g1212995 BLASTN 388 1e−48 80 1465 69 700219739H1 SATMON011 g1212995 BLASTN 443 1e−48 81 1466 69 700620336H1 SATMON034 g1212995 BLASTN 621 1e−48 88 1467 69 700264630H1 SATMON017 g1212995 BLASTN 377 1e−47 80 1468 69 700439242H1 SATMON026 g1212995 BLASTN 648 1e−47 83 1469 69 700259658H1 SATMON017 g1212995 BLASTN 511 1e−45 79 1470 69 700263521H1 SATMON017 g1212995 BLASTN 461 1e−44 79 1471 69 700261387H1 SATMON017 g1212995 BLASTN 461 1e−44 80 1472 69 700439277H1 SATMON026 g1212995 BLASTN 461 1e−43 84 1473 69 700452839H1 SATMON028 g1212995 BLASTN 544 1e−43 77 1474 69 700220236H1 SATMON011 g1212995 BLASTN 448 1e−40 84 1475 69 700472602H1 SATMON025 g1212995 BLASTN 254 1e−38 81 1476 69 700266424H1 SATMON017 g1212995 BLASTN 499 1e−37 80 1477 69 700449187H1 SATMON028 g1212995 BLASTN 540 1e−36 81 1478 69 700202731H1 SATMON003 g1212995 BLASTN 543 1e−36 79 1479 69 700156144H2 SATMON007 g1212995 BLASTN 441 1e−35 76 1480 69 700442679H1 SATMON026 g1212995 BLASTN 533 1e−35 80 1481 69 700449879H2 SATMON028 g1212995 BLASTN 535 1e−35 81 1482 69 700266832H1 SATMON017 g1212995 BLASTN 346 1e−34 77 1483 69 700332389H1 SATMON019 g1212995 BLASTN 382 1e−34 84 1484 69 700804202H1 SATMON036 g1212995 BLASTN 436 1e−34 76 1485 69 700151037H1 SATMON007 g1212995 BLASTN 443 1e−34 79 1486 69 700802810H1 SATMON036 g1212995 BLASTN 525 1e−34 85 1487 69 700455879H1 SATMON029 g1212995 BLASTN 448 1e−32 72 1488 69 700427769H1 SATMONN01 g1212995 BLASTN 481 1e−31 81 1489 69 700464626H1 SATMON025 g1212995 BLASTN 388 1e−30 76 1490 69 700439228H1 SATMON026 g1212995 BLASTN 470 1e−30 77 1491 69 700256847H1 SATMON017 g1212995 BLASTN 264 1e−29 85 1492 69 700204881H1 SATMON003 g1212995 BLASTN 430 1e−29 81 1493 69 700076032H1 SATMON007 g1212995 BLASTN 218 1e−26 72 1494 69 700426342H1 SATMONN01 g1212995 BLASTN 443 1e−26 79 1495 69 700209062H1 SATMON016 g1212995 BLASTN 279 1e−24 80 1496 69 700076988H1 SATMON007 g1212995 BLASTN 337 1e−24 83 1497 69 700349778H1 SATMON023 g1212995 BLASTN 406 1e−24 81 1498 69 700261886H1 SATMON017 g1212995 BLASTN 287 1e−15 80 1499 69 700426642H1 SATMONN01 g1388021 BLASTX 161 1e−14 76 1500 69 700155195H1 SATMON007 g1212995 BLASTN 155 1e−10 81 1501 69 700211992H1 SATMON016 g1212996 BLASTX 118 1e−9 85 1502 −L1485255 LIB148-053- LIB148 g1212995 BLASTN 691 1e−48 80 Q1-E1-E12 1503 −L30663959 LIB3066-015- LIB3066 g218000 BLASTN 251 1e−9 65 Q1-K1-F12 1504 19537 LIB3066-025- LIB3066 g1212995 BLASTN 1001 1e−74 79 Q1-K1-E5 1505 69 LIB3059-023- LIB3059 g1212995 BLASTN 1301 1e−133 89 Q1-K1-C8 1506 69 LIB3078-022- LIB3078 g1212995 BLASTN 1656 1e−129 86 Q1-K1-C1 1507 69 LIB3059-037- LIB3059 g1212995 BLASTN 1646 1e−128 86 Q1-K1-H5 1508 69 LIB3061-030- LIB3061 g1212995 BLASTN 1493 1e−124 86 Q1-K1-A12 1509 69 LIB3061-023- LIB3061 g1212995 BLASTN 1598 1e−124 86 Q1-K1-A1 1510 69 LIB3079-001- LIB3079 g1212995 BLASTN 1600 1e−124 83 Q1-K1-D12 1511 69 LIB189-028- LIB189 g1212995 BLASTN 1583 1e−123 87 Q1-E1-E3 1512 69 LIB3067-017- LIB3067 g1212995 BLASTN 1364 1e−120 88 Q1-K1-D9 1513 69 LIB3068-007- LIB3068 g1212995 BLASTN 1501 1e−116 85 Q1-K1-F9 1514 69 LIB3069-025- LIB3069 g1212995 BLASTN 1487 1e−115 85 Q1-K1-E9 1515 69 LIB3069-026- LIB3069 g1212995 BLASTN 1453 1e−112 85 Q1-K1-E11 1516 69 LIB3066-006- LIB3066 g1212995 BLASTN 1077 1e−107 83 Q1-K1-G12 1517 69 LIB3067-027- LIB3067 g1212995 BLASTN 1401 1e−107 86 Q1-K1-D12 1518 69 LIB189-010- LIB189 g1212995 BLASTN 1368 1e−105 85 Q1-E1-H10 1519 69 LIB3066-015- LIB3066 g1212995 BLASTN 1289 1e−104 82 Q1-K1-G12 1520 69 LIB3061-016- LIB3061 g1212995 BLASTN 1180 1e−102 84 Q1-K1-G11 1521 69 LIB3059-032- LIB3059 g1212995 BLASTN 1334 1e−102 87 Q1-K1-G11 1522 69 LIB3067-059- LIB3067 g1212995 BLASTN 1090 1e−100 85 Q1-K1-G12 1523 69 LIB3061-049- LIB3061 g1212995 BLASTN 1223 1e−98 79 Q1-K1-C8 1524 69 LIB3062-044- LIB3062 g1212995 BLASTN 1259 1e−96 83 Q1-K1-F2 1525 69 LIB3061-010- LIB3061 g1212995 BLASTN 1180 1e−95 84 Q1-K1-F5 1526 69 LIB3067-018- LIB3067 g1212995 BLASTN 1127 1e−89 82 Q1-K1-A12 1527 69 LIB3067-030- LIB3067 g1212995 BLASTN 1171 1e−88 83 Q1-K1-F4 1528 69 LIB3062-021- LIB3062 g1212995 BLASTN 1138 1e−86 87 Q1-K1-F10 1529 69 LIB3061-034- LIB3061 g1212995 BLASTN 1148 1e−86 85 Q1-K1-D8 1530 69 LIB3066-049- LIB3066 g1212995 BLASTN 1134 1e−85 83 Q1-K1-C1 1531 69 LIB3078-002- LIB3078 g1212995 BLASTN 859 1e−77 86 Q1-K1-F5 1532 69 LIB84-011- LIB84 g1212995 BLASTN 1020 1e−76 83 Q1-E1-G9 1533 69 LIB3067-043- LIB3067 g1212995 BLASTN 574 1e−59 77 Q1-K1-D2 1534 69 LIB189-003- LIB189 g1212995 BLASTN 247 1e−40 77 Q1-E1-G5 1535 69 LIB3062-008- LIB3062 g1212995 BLASTN 576 1e−37 63 Q1-K1-E6 1536 69 LIB189-016- LIB189 g1212996 BLASTX 156 1e−30 78 Q1-E1-H7 1537 69 LIB3067-007- LIB3067 g1212996 BLASTX 145 1e−28 82 Q1-K1-G4 SOYBEAN TRIOSE PHOSPHATE ISOMERASE 1538 −700743237 700743237H1 SOYMON012 g407525 BLASTX 173 1e−17 91 1539 −700977730 700977730H1 SOYMON009 g602589 BLASTN 373 1e−20 71 1540 −701056176 701056176H1 SOYMON032 g806311 BLASTN 752 1e−53 74 1541 −701110172 701110172H1 SOYMON036 g806311 BLASTN 801 1e−57 78 1542 10244 700995141H1 SOYMON011 g806311 BLASTN 470 1e−30 87 1543 10244 701124548H1 SOYMON037 g806311 BLASTN 490 1e−30 88 1544 10244 700739771H1 SOYMON012 g806311 BLASTN 329 1e−16 77 1545 10244 700999820H1 SOYMON018 g806312 BLASTX 147 1e−13 84 1546 10244 701119858H1 SOYMON037 g806312 BLASTX 118 1e−9 72 1547 10535 700988684H1 SOYMON009 g806311 BLASTN 905 1e−66 79 1548 10535 700902425H1 SOYMON027 g806311 BLASTN 872 1e−63 80 1549 1357 701069004H1 SOYMON034 g806311 BLASTN 832 1e−60 81 1550 1357 701151554H1 SOYMON031 g806311 BLASTN 568 1e−38 82 1551 1357 700659936H1 SOYMON004 g806311 BLASTN 545 1e−36 79 1552 16 700680927H1 SOYMON008 g256119 BLASTN 1020 1e−81 78 1553 16 700656871H1 SOYMON004 g256119 BLASTN 903 1e−66 81 1554 16 701124364H1 SOYMON037 g256119 BLASTN 872 1e−64 80 1555 16 701134707H2 SOYMON038 g256119 BLASTN 874 1e−64 81 1556 16 700673750H1 SOYMON007 g256119 BLASTN 781 1e−60 81 1557 16 701123269H1 SOYMON037 g602589 BLASTN 819 1e−59 78 1558 16 701004846H1 SOYMON019 g256119 BLASTN 801 1e−58 80 1559 16 700993362H1 SOYMON011 g256119 BLASTN 808 1e−58 80 1560 16 701005445H1 SOYMON019 g256119 BLASTN 630 1e−56 78 1561 16 701134327H1 SOYMON038 g602589 BLASTN 782 1e−56 79 1562 16 701148169H1 SOYMON031 g602589 BLASTN 574 1e−51 76 1563 16 701153410H1 SOYMON031 g602589 BLASTN 451 1e−50 80 1564 16 700830168H1 SOYMON019 g256119 BLASTN 705 1e−50 77 1565 16 701120627H1 SOYMON037 g602589 BLASTN 715 1e−50 78 1566 16 700975358H1 SOYMON009 g602589 BLASTN 628 1e−49 77 1567 16 700755979H1 SOYMON014 g602589 BLASTN 697 1e−49 79 1568 16 701131374H1 SOYMON038 g602589 BLASTN 703 1e−49 79 1569 16 700994166H1 SOYMON011 g602589 BLASTN 513 1e−47 77 1570 16 701138038H1 SOYMON038 g602589 BLASTN 672 1e−47 77 1571 16 700974248H1 SOYMON005 g602589 BLASTN 658 1e−46 77 1572 16 700655832H1 SOYMON004 g602589 BLASTN 664 1e−46 78 1573 16 700758320H1 SOYMON015 g602589 BLASTN 409 1e−45 80 1574 16 701064709H1 SOYMON034 g602589 BLASTN 477 1e−45 78 1575 16 701138504H1 SOYMON038 g602589 BLASTN 591 1e−45 76 1576 16 700980284H1 SOYMON009 g602589 BLASTN 652 1e−45 79 1577 16 701133585H2 SOYMON038 g602589 BLASTN 634 1e−44 78 1578 16 700674706H1 SOYMON007 g602589 BLASTN 634 1e−44 78 1579 16 700964927H1 SOYMON022 g602589 BLASTN 639 1e−44 78 1580 16 700830923H1 SOYMON019 g602589 BLASTN 626 1e−43 76 1581 16 700662845H1 SOYMON005 g602589 BLASTN 617 1e−42 76 1582 16 701133824H1 SOYMON038 g602589 BLASTN 619 1e−42 78 1583 16 700848913H1 SOYMON021 g602589 BLASTN 603 1e−41 77 1584 16 701005984H1 SOYMON019 g602589 BLASTN 604 1e−41 78 1585 16 701140769H1 SOYMON038 g602589 BLASTN 605 1e−41 76 1586 16 700753357H1 SOYMON014 g602589 BLASTN 328 1e−40 78 1587 16 701056336H1 SOYMON032 g602589 BLASTN 344 1e−40 77 1588 16 700895411H1 SOYMON027 g602589 BLASTN 593 1e−40 78 1589 16 701060188H1 SOYMON033 g602589 BLASTN 277 1e−39 80 1590 16 700739461H1 SOYMON012 g602589 BLASTN 573 1e−39 77 1591 16 700941104H1 SOYMON024 g602589 BLASTN 579 1e−39 79 1592 16 700732960H1 SOYMON010 g602589 BLASTN 581 1e−39 78 1593 16 700686476H1 SOYMON008 g602589 BLASTN 583 1e−39 79 1594 16 701054231H1 SOYMON032 g602589 BLASTN 583 1e−39 77 1595 16 700671690H1 SOYMON006 g602589 BLASTN 566 1e−38 77 1596 16 700941174H1 SOYMON024 g602589 BLASTN 569 1e−38 78 1597 16 701125091H1 SOYMON037 g256119 BLASTN 358 1e−37 74 1598 16 700989827H1 SOYMON011 g602589 BLASTN 555 1e−37 78 1599 16 700835006H1 SOYMON019 g602589 BLASTN 555 1e−37 75 1600 16 700834847H1 SOYMON019 g602589 BLASTN 559 1e−37 78 1601 16 700953411H1 SOYMON022 g602589 BLASTN 314 1e−36 80 1602 16 700869222H1 SOYMON016 g602589 BLASTN 541 1e−36 78 1603 16 700850633H1 SOYMON023 g602589 BLASTN 544 1e−36 78 1604 16 700890283H1 SOYMON024 g602589 BLASTN 310 1e−35 80 1605 16 700727079H1 SOYMON009 g414549 BLASTN 358 1e−35 73 1606 16 700892544H1 SOYMON024 g602589 BLASTN 486 1e−35 78 1607 16 700869230H1 SOYMON016 g602589 BLASTN 528 1e−35 78 1608 16 700993034H1 SOYMON011 g602589 BLASTN 518 1e−34 75 1609 16 700975553H1 SOYMON009 g414549 BLASTN 524 1e−34 79 1610 16 700651326H1 SOYMON003 g602589 BLASTN 356 1e−33 80 1611 16 701215308H1 SOYMON035 g414549 BLASTN 450 1e−33 75 1612 16 700654480H1 SOYMON004 g414549 BLASTN 511 1e−33 80 1613 16 701045128H1 SOYMON032 g414549 BLASTN 512 1e−33 78 1614 16 701060759H1 SOYMON033 g414549 BLASTN 513 1e−33 80 1615 16 700741652H1 SOYMON012 g602589 BLASTN 493 1e−32 79 1616 16 700675469H1 SOYMON007 g602589 BLASTN 494 1e−32 78 1617 16 700657787H1 SOYMON004 g414549 BLASTN 495 1e−32 79 1618 16 701009957H2 SOYMON019 g414549 BLASTN 495 1e−32 80 1619 16 700983693H1 SOYMON009 g414549 BLASTN 495 1e−32 80 1620 16 701156784H1 SOYMON031 g602589 BLASTN 495 1e−32 78 1621 16 700893935H1 SOYMON024 g602589 BLASTN 481 1e−31 79 1622 16 701144619H1 SOYMON031 g414549 BLASTN 485 1e−31 78 1623 16 701148851H1 SOYMON031 g602589 BLASTN 487 1e−31 79 1624 16 701058218H1 SOYMON033 g602589 BLASTN 495 1e−31 78 1625 16 700975165H1 SOYMON009 g414549 BLASTN 466 1e−30 80 1626 16 701100165H1 SOYMON028 g602589 BLASTN 485 1e−30 79 1627 16 701150241H1 SOYMON031 g602589 BLASTN 455 1e−29 79 1628 16 701098308H1 SOYMON028 g414549 BLASTN 460 1e−29 79 1629 16 701150440H1 SOYMON031 g602589 BLASTN 462 1e−29 78 1630 16 700685125H1 SOYMON008 g414549 BLASTN 471 1e−29 81 1631 16 701061565H1 SOYMON033 g414549 BLASTN 471 1e−29 81 1632 16 700991418H1 SOYMON011 g602589 BLASTN 394 1e−28 68 1633 16 701156156H1 SOYMON031 g602589 BLASTN 456 1e−28 78 1634 16 701007231H2 SOYMON019 g602589 BLASTN 461 1e−28 79 1635 16 700829667H1 SOYMON019 g414549 BLASTN 333 1e−27 73 1636 16 701156033H1 SOYMON031 g602589 BLASTN 432 1e−27 78 1637 16 701014293H1 SOYMON019 g414549 BLASTN 446 1e−27 77 1638 16 701152138H1 SOYMON031 g414549 BLASTN 450 1e−27 81 1639 16 700945665H1 SOYMON024 g414549 BLASTN 450 1e−27 81 1640 16 701001407H1 SOYMON018 g169820 BLASTN 219 1e−26 72 1641 16 700983185H1 SOYMON009 g414549 BLASTN 435 1e−26 72 1642 16 700752364H1 SOYMON014 g414549 BLASTN 441 1e−26 76 1643 16 700992409H1 SOYMON011 g414549 BLASTN 427 1e−25 75 1644 16 701109396H1 SOYMON036 g414549 BLASTN 420 1e−24 76 1645 16 701151402H1 SOYMON031 g556171 BLASTX 151 1e−23 85 1646 16 701149617H1 SOYMON031 g556171 BLASTX 158 1e−23 86 1647 16 700747310H1 SOYMON013 g414549 BLASTN 406 1e−23 73 1648 16 701139569H1 SOYMON038 g556171 BLASTX 191 1e−22 84 1649 16 701213275H1 SOYMON035 g602589 BLASTN 255 1e−22 80 1650 16 701157185H1 SOYMON031 g556171 BLASTX 197 1e−20 90 1651 16 700655520H1 SOYMON004 g556171 BLASTX 166 1e−19 86 1652 16 701010779H1 SOYMON019 g556171 BLASTX 173 1e−19 64 1653 16 701044104H1 SOYMON032 g556171 BLASTX 188 1e−19 89 1654 16 700867605H1 SOYMON016 g556171 BLASTX 160 1e−17 70 1655 16 701058593H1 SOYMON033 g168647 BLASTX 169 1e−16 94 1656 16 701070286H1 SOYMON034 g168647 BLASTX 164 1e−15 91 1657 16 700877219H1 SOYMON018 g168647 BLASTX 154 1e−14 93 1658 16 700876790H1 SOYMON018 g168647 BLASTX 154 1e−14 93 1659 16 700877212H1 SOYMON018 g168647 BLASTX 154 1e−14 93 1660 16 700760847H1 SOYMON015 g556171 BLASTX 138 1e−13 86 1661 16 700893711H1 SOYMON024 g168647 BLASTX 140 1e−13 82 1662 16 700557532H1 SOYMON001 g256120 BLASTX 115 1e−12 88 1663 16 700793802H1 SOYMON017 g556171 BLASTX 138 1e−12 93 1664 16 700659725H1 SOYMON004 g556171 BLASTX 144 1e−12 47 1665 16 701044545H1 SOYMON032 g556171 BLASTX 144 1e−12 92 1666 16 701037485H1 SOYMON029 g556171 BLASTX 135 1e−11 96 1667 16 700683524H1 SOYMON008 g168647 BLASTX 136 1e−11 90 1668 16 700876711H1 SOYMON018 g168647 BLASTX 109 1e−10 85 1669 16 701155437H1 SOYMON031 g556171 BLASTX 130 1e−10 92 1670 28599 700997892H1 SOYMON018 g806311 BLASTN 834 1e−60 78 1671 31 701053174H1 SOYMON032 g806311 BLASTN 572 1e−37 73 1672 31 700754467H1 SOYMON014 g806312 BLASTX 145 1e−21 66 1673 31 701107430H1 SOYMON036 g806312 BLASTX 199 1e−20 63 1674 31 700985855H1 SOYMON009 g806312 BLASTX 145 1e−18 64 1675 31 701038167H1 SOYMON029 g806312 BLASTX 179 1e−17 61 1676 31 700670393H1 SOYMON006 g806312 BLASTX 167 1e−16 78 1677 31 700559280H1 SOYMON001 g609262 BLASTX 164 1e−15 69 1678 31 700793048H1 SOYMON017 g806312 BLASTX 97 1e−12 60 1679 31 700993683H1 SOYMON011 g806312 BLASTX 103 1e−11 60 1680 31 700663233H1 SOYMON005 g806312 BLASTX 130 1e−11 56 1681 31 700908079H1 SOYMON022 g806312 BLASTX 103 1e−10 60 1682 31 701043447H1 SOYMON029 g609262 BLASTX 126 1e−10 84 1683 31 700740188H1 SOYMON012 g806312 BLASTX 103 1e−8 60 1684 7466 700742922H1 SOYMON012 g806311 BLASTN 435 1e−27 76 1685 7466 700606255H1 SOYMON008 g806312 BLASTX 117 1e−17 80 1686 16 LIB3053-005- LIB3053 g602589 BLASTN 1000 1e−74 77 Q1-N1-F9 1687 16 LIB3039-035- LIB3039 g602589 BLASTN 979 1e−72 78 Q1-E1-C5 1688 16 LIB3039-031- LIB3039 g256119 BLASTN 911 1e−71 80 Q1-E1-A8 1689 16 LIB3030-003- LIB3030 g602589 BLASTN 949 1e−70 78 Q1-B1-C9 1690 16 LIB3039-023- LIB3039 g602589 BLASTN 913 1e−67 78 Q1-E1-H12 1691 16 LIB3039-047- LIB3039 g602589 BLASTN 566 1e−65 75 Q1-E1-D8 1692 16 LIB3039-052- LIB3039 g602589 BLASTN 890 1e−65 77 Q1-E1-D6 1693 16 LIB3039-051- LIB3039 g602589 BLASTN 855 1e−62 78 Q1-E1-A1 1694 16 LIB3049-009- LIB3049 g602589 BLASTN 783 1e−56 78 Q1-E1-G5 1695 16 LIB3039-009- LIB3039 g602589 BLASTN 805 1e−56 78 Q1-E1-C1 1696 16 LIB3055-006- LIB3055 g256119 BLASTN 481 1e−54 78 Q1-N1-H3 1697 16 LIB3055-013- LIB3055 g256119 BLASTN 769 1e−54 79 Q1-N1-C3 1698 16 LIB3049-034- LIB3049 g602589 BLASTN 626 1e−51 76 Q1-E1-A2 1699 16 LIB3049-022- LIB3049 g602589 BLASTN 519 1e−43 78 Q1-E1-F9 1700 16 LIB3049-030- LIB3049 g602589 BLASTN 572 1e−38 77 Q1-E1-C7 1701 16 LIB3040-035- LIB3040 g556171 BLASTX 175 1e−33 82 Q1-E1-C5 1702 16 LIB3040-005- LIB3040 g169820 BLASTN 324 1e−33 76 Q1-E1-H8 1703 16 LIB3028-025- LIB3028 g602589 BLASTN 464 1e−33 78 Q1-B1-D1 1704 16 LIB3039-022- LIB3039 g602589 BLASTN 357 1e−32 73 Q1-E1-D5 1705 16 LIB3052-001- LIB3052 G414549 BLASTN 327 1e−29 73 Q1-B1-C5 1706 28599 LIB3039-047- LIB3039 G806311 BLASTN 1183 1e−94 81 Q1-E1-D9 1707 28599 LIB3039-048- LIB3039 G806311 BLASTN 1007 1e−92 81 Q1-E1-D12 SOYBEAN FRUCTOSE 1,6-BISPHOSPHATE ALDOLASE 1708 00565253 700565253H1 SOYMON002 G3021337 BLASTN 352 1e−39 76 1709 −700865276 700865276H1 SOYMON016 G3021337 BLASTN 629 1e−43 76 1710 −700873022 700873022H1 SOYMON018 G3696 BLASTX 211 1e−26 70 1711 −700943855 700943855H1 SOYMON024 G20204 BLASTX 202 1e−20 86 1712 −700974965 700974965H1 SOYMON005 g3021337 BLASTN 259 1e−10 84 1713 −701039850 701039850H1 SOYMON029 g22632 BLASTN 408 1e−23 76 1714 −701206840 701206840H1 SOYMON035 g3021338 BLASTX 151 1e−13 83 1715 11792 700654881H1 SOYMON004 g20204 BLASTX 150 1e−13 76 1716 11792 700746016H1 SOYMON013 g3021337 BLASTN 284 1e−12 67 1717 12314 701037190H1 SOYMON029 g3021337 BLASTN 634 1e−44 78 1718 12314 701042664H1 SOYMON029 g3021338 BLASTX 197 1e−20 66 1719 16 700651596H1 SOYMON003 g3021337 BLASTN 1101 1e−83 86 1720 16 700750439H1 SOYMON013 g3021337 BLASTN 1078 1e−81 86 1721 16 700649475H1 SOYMON003 g3021337 BLASTN 1082 1e−81 84 1722 16 700652995H1 SOYMON003 g3021337 BLASTN 1084 1e−81 82 1723 16 700981967H1 SOYMON009 g3021337 BLASTN 1071 1e−80 85 1724 16 700863243H1 SOYMON023 g3021337 BLASTN 1044 1e−78 86 1725 16 700558625H1 SOYMON001 g3021337 BLASTN 1041 1e−77 84 1726 16 700564806H1 SOYMON002 g3021337 BLASTN 1021 1e−76 80 1727 16 700746368H1 SOYMON013 g3021337 BLASTN 897 1e−75 86 1728 16 700960290H1 SOYMON022 g3021337 BLASTN 1009 1e−75 87 1729 16 701055132H1 SOYMON032 g3021337 BLASTN 1011 1e−75 86 1730 16 701056109H1 SOYMON032 g3021337 BLASTN 1012 1e−75 84 1731 16 701119884H1 SOYMON037 g3021337 BLASTN 1014 1e−75 87 1732 16 700898149H1 SOYMON027 g3021337 BLASTN 1015 1e−75 86 1733 16 700661436H1 SOYMON005 g3021337 BLASTN 596 1e−74 83 1734 16 701042223H1 SOYMON029 g3021337 BLASTN 997 1e−74 84 1735 16 700676004H1 SOYMON007 g3021337 BLASTN 984 1e−73 85 1736 16 700747718H1 SOYMON013 g3021337 BLASTN 988 1e−73 87 1737 16 700751133H1 SOYMON014 g3021337 BLASTN 989 1e−73 86 1738 16 701215247H1 SOYMON035 g3021337 BLASTN 989 1e−73 84 1739 16 700652484H1 SOYMON003 g3021337 BLASTN 910 1e−72 85 1740 16 700869785H1 SOYMON016 g3021337 BLASTN 970 1e−72 87 1741 16 700981960H1 SOYMON009 g3021337 BLASTN 970 1e−72 87 1742 16 700969335H1 SOYMON005 g3021337 BLASTN 972 1e−72 82 1743 16 700854174H1 SOYMON023 g3021337 BLASTN 965 1e−71 84 1744 16 700761638H1 SOYMON015 g3021337 BLASTN 966 1e−71 86 1745 16 700984860H1 SOYMON009 g3021337 BLASTN 967 1e−71 84 1746 16 701005716H1 SOYMON019 g3021337 BLASTN 967 1e−71 83 1747 16 700941053H1 SOYMON024 g3021337 BLASTN 968 1e−71 86 1748 16 700561358H1 SOYMON002 g3021337 BLASTN 968 1e−71 82 1749 16 700564906H1 SOYMON002 g3021337 BLASTN 562 1e−70 82 1750 16 700833951H1 SOYMON019 g3021337 BLASTN 954 1e−70 88 1751 16 701117626H1 SOYMON037 g3021337 BLASTN 957 1e−70 85 1752 16 700729103H1 SOYMON009 g3021337 BLASTN 535 1e−69 86 1753 16 700670615H1 SOYMON006 g3021337 BLASTN 936 1e−69 83 1754 16 701053635H1 SOYMON032 g3021337 BLASTN 941 1e−69 84 1755 16 700982280H1 SOYMON009 g3021337 BLASTN 923 1e−68 82 1756 16 701119874H1 SOYMON037 g3021337 BLASTN 925 1e−68 88 1757 16 700758937H1 SOYMON015 g3021337 BLASTN 926 1e−68 87 1758 16 701214027H1 SOYMON035 g3021337 BLASTN 928 1e−68 82 1759 16 700972858H1 SOYMON005 g3021337 BLASTN 929 1e−68 84 1760 16 701099780H1 SOYMON028 g3021337 BLASTN 930 1e−68 85 1761 16 700829560H1 SOYMON019 g3021337 BLASTN 932 1e−68 85 1762 16 700971973H1 SOYMON005 g3021337 BLASTN 576 1e−67 85 1763 16 701142336H1 SOYMON038 g3021337 BLASTN 750 1e−67 81 1764 16 701132605H1 SOYMON038 g3021337 BLASTN 759 1e−67 85 1765 16 700969222H1 SOYMON005 g3021337 BLASTN 913 1e−67 84 1766 16 700670956H1 SOYMON006 g3021337 BLASTN 920 1e−67 84 1767 16 700895725H1 SOYMON027 g3021337 BLASTN 921 1e−67 84 1768 16 701013771H1 SOYMON019 g3021337 BLASTN 921 1e−67 81 1769 16 701055481H1 SOYMON032 g3021337 BLASTN 654 1e−66 80 1770 16 700753940H1 SOYMON014 g3021337 BLASTN 899 1e−66 84 1771 16 700974141H1 SOYMON005 g3021337 BLASIN 900 1e−66 81 1772 16 700562408H1 SOYMON002 g3021337 BLASTN 902 1e−66 82 1773 16 700685292H1 SOYMON008 g3021337 BLASTN 903 1e−66 83 1774 16 700985157H1 SOYMON009 g3021337 BLASTN 907 1e−66 82 1775 16 701038194H1 SOYMON029 g3021337 BLASTN 907 1e−66 82 1776 16 700986633H1 SOYMON009 g3021337 BLASTN 908 1e−66 83 1777 16 700564282H1 SOYMON002 g3021337 BLASTN 517 1e−65 83 1778 16 700733754H1 SOYMON010 g3021337 BLASTN 680 1e−65 84 1779 16 700988179H1 SOYMON009 g3021337 BLASTN 887 1e−65 82 1780 16 700555591H1 SOYMON001 g3021337 BLASTN 887 1e−65 82 1781 16 701206717H1 SOYMON035 g3021337 BLASTN 888 1e−65 81 1782 16 700968494H1 SOYMON036 g3021337 BLASTN 889 1e−65 86 1783 16 700906271H1 SOYMON022 g3021337 BLASTN 894 1e−65 82 1784 16 700677674H1 SOYMON007 g3021337 BLASTN 894 1e−65 83 1785 16 700970391H1 SOYMON005 g3021337 BLASTN 896 1e−65 83 1786 16 700753641H1 SOYMON014 g3021337 BLASTN 897 1e−65 82 1787 16 700646593H1 SOYMON014 g3021337 BLASTN 468 1e−64 80 1788 16 700565615H1 SOYMON002 g3021337 BLASTN 667 1e−64 80 1789 16 700746523H1 SOYMON013 g3021337 BLASTN 744 1e−64 83 1790 16 700899019H1 SOYMON027 g3021337 BLASTN 875 1e−64 83 1791 16 701127167H1 SOYMON037 g3021337 BLASTN 876 1e−64 84 1792 16 701131053H1 SOYMON038 g3021337 BLASTN 879 1e−64 84 1793 16 700670980H1 SOYMON006 g3021337 BLASTN 881 1e−64 83 1794 16 701055811H1 SOYMON032 g3021337 BLASTN 881 1e−64 85 1795 16 700900103H1 SOYMON027 g3021337 BLASTN 882 1e−64 83 1796 16 700975609H1 SOYMON009 g3021337 BLASTN 882 1e−64 84 1797 16 701102865H1 SOYMON028 g3021337 BLASTN 883 1e−64 85 1798 16 701145255H1 SOYMON031 g3021337 BLASTN 509 1e−63 80 1799 16 701210875H1 SOYMON035 g3021337 BLASTN 616 1e−63 84 1800 16 700646664H1 SOYMON014 g3021337 BLASTN 862 1e−63 85 1801 16 700897337H1 SOYMON027 g3021337 BLASTN 865 1e−63 86 1802 16 700736783H1 SOYMON010 g3021337 BLASTN 867 1e−63 83 1803 16 701059586H1 SOYMON033 g3021337 BLASTN 869 1e−63 81 1804 16 701127063H1 SOYMON037 g3021337 BLASTN 412 1e−62 84 1805 16 700556614H1 SOYMON001 g3021337 BLASTN 475 1e−62 86 1806 16 700672681H1 SOYMON006 g3021337 BLASTN 818 1e−62 82 1807 16 700727057H1 SOYMON009 g3021337 BLASTN 850 1e−62 82 1808 16 701042141H1 SOYMON029 g3021337 BLASTN 851 1e−62 83 1809 16 700561860H1 SOYMON002 g3021337 BLASTN 854 1e−62 81 1810 16 700677460H1 SOYMON007 g3021337 BLASTN 855 1e−62 83 1811 16 700971671H1 SOYMON005 g3021337 BLASTN 856 1e−62 81 1812 16 700749578H1 SOYMON013 g3021337 BLASTN 856 1e−62 81 1813 16 700672288H1 SOYMON006 g3021337 BLASTN 860 1e−62 81 1814 16 701068481H1 SOYMON034 g3021337 BLASTN 861 1e−62 81 1815 16 700729913H1 SOYMON009 g3021337 BLASTN 661 1e−61 79 1816 16 700739449H1 SOYMON012 g3021337 BLASTN 724 1e−61 85 1817 16 700830902H1 SOYMON019 g3021337 BLASTN 814 1e−61 83 1818 16 700895304H1 SOYMON027 g3021337 BLASTN 840 1e−61 82 1819 16 700605676H2 SOYMON005 g3021337 BLASTN 842 1e−61 84 1820 16 700677453H1 SOYMON007 g3021337 BLASTN 843 1e−61 83 1821 16 700983108H1 SOYMON009 g3021337 BLASTN 843 1e−61 81 1822 16 700889170H1 SOYMON024 g3021337 BLASTN 845 1e−61 86 1823 16 701004956H1 SOYMON019 g3021337 BLASTN 849 1e−61 82 1824 16 700958213H1 SOYMON022 g3021337 BLASTN 849 1e−61 82 1825 16 701129305H1 SOYMON037 g3021337 BLASTN 659 1e−60 85 1826 16 701014446H1 SOYMON019 g3021337 BLASTN 669 1e−60 85 1827 16 700832047H1 SOYMON019 g3021337 BLASTN 738 1e−60 83 1828 16 700669966H1 SOYMON006 g3021337 BLASTN 829 1e−60 82 1829 16 700758028H1 SOYMON015 g3021337 BLASTN 829 1e−60 81 1830 16 700659491H1 SOYMON004 g3021337 BLASTN 829 1e−60 83 1831 16 701003560H1 SOYMON019 g3021337 BLASTN 829 1e−60 82 1832 16 701060964H1 SOYMON033 g3021337 BLASTN 833 1e−60 81 1833 16 700548284H1 SOYMON002 g3021337 BLASTN 834 1e−60 82 1834 16 700894957H1 SOYMON024 g3021337 BLASTN 837 1e−60 81 1835 16 700646551H1 SOYMON014 g3021337 BLASTN 479 1e−59 83 1836 16 700967633H1 SOYMON032 g3021337 BLASTN 530 1e−59 81 1837 16 700754430H1 SOYMON014 g3021337 BLASTN 654 1e−59 85 1838 16 700865919H1 SOYMON016 g3021337 BLASTN 814 1e−59 81 1839 16 700980426H1 SOYMON009 g3021337 BLASTN 815 1e−59 80 1840 16 701048203H1 SOYMON032 g3021337 BLASTN 816 1e−59 81 1841 16 700846414H1 SOYMON021 g3021337 BLASTN 819 1e−59 81 1842 16 700851608H1 SOYMON023 g3021337 BLASTN 822 1e−59 81 1843 16 700970160H1 SOYMON005 g3021337 BLASTN 822 1e−59 82 1844 16 700834462H1 SOYMON019 g3021337 BLASTN 823 1e−59 81 1845 16 701206312H1 SOYMON035 g3021337 BLASTN 823 1e−59 85 1846 16 700562478H1 SOYMON002 g3021337 BLASTN 487 1e−58 84 1847 16 700788114H1 SOYMON011 g3021337 BLASTN 751 1e−58 83 1848 16 700753792H1 SOYMON014 g3021337 BLASTN 804 1e−58 84 1849 16 700837427H1 SOYMON020 g3021337 BLASTN 805 1e−58 86 1850 16 700753668H1 SOYMON014 g3021337 BLASTN 806 1e−58 85 1851 16 700667315H1 SOYMON006 g3021337 BLASTN 809 1e−58 81 1852 16 700808315H1 SOYMON024 g3021337 BLASTN 558 1e−57 80 1853 16 700670207H1 SOYMON006 g3021337 BLASTN 791 1e−57 87 1854 16 700849886H1 SOYMON021 g3021337 BLASTN 791 1e−57 83 1855 16 700839033H1 SOYMON020 g3021337 BLASTN 791 1e−57 81 1856 16 700751117H1 SOYMON014 g3021337 BLASTN 799 1e−57 86 1857 16 700851803H1 SOYMON023 g3021337 BLASTN 799 1e−57 86 1858 16 700669164H1 SOYMON006 g3021337 BLASTN 800 1e−57 80 1859 16 700548285H1 SOYMON002 g3021337 BLASTN 801 1e−57 85 1860 16 701065620H1 SOYMON034 g3021337 BLASTN 426 1e−56 82 1861 16 700727996H1 SOYMON009 g3021337 BLASTN 468 1e−56 79 1862 16 700869176H1 SOYMON016 g3021337 BLASTN 786 1e−56 85 1863 16 700973141H1 SOYMON005 g3021337 BLASTN 440 1e−55 79 1864 16 700969555H1 SOYMON005 g3021337 BLASTN 448 1e−55 81 1865 16 700866138H1 SOYMON016 g3021337 BLASTN 641 1e−55 86 1866 16 700904813H1 SOYMON022 g3021337 BLASTN 699 1e−55 85 1867 16 700894146H1 SOYMON024 g3021337 BLASTN 773 1e−55 86 1868 16 700669945H1 SOYMON006 g3021337 BLASTN 773 1e−55 86 1869 16 701060489H1 SOYMON033 g3021337 BLASTN 664 1e−54 85 1870 16 701125675H1 SOYMON037 g3021337 BLASTN 721 1e−54 85 1871 16 700754750H1 SOYMON014 g3021337 BLASTN 722 1e−54 86 1872 16 701142770H1 SOYMON038 g3021337 BLASTN 755 1e−54 88 1873 16 700731095H1 SOYMON009 g3021337 BLASTN 755 1e−54 87 1874 16 700667966H1 SOYMON006 g3021337 BLASTN 756 1e−54 84 1875 16 700673606H1 SOYMON007 g3021337 BLASTN 760 1e−54 83 1876 16 700605289H2 SOYMON003 g3021337 BLASTN 763 1e−54 84 1877 16 700965253H1 SOYMON022 g3021337 BLASTN 763 1e−54 86 1878 16 700732985H1 SOYMON010 g3021337 BLASTN 765 1e−54 87 1879 16 700986523H1 SOYMON009 g3021337 BLASTN 474 1e−53 85 1880 16 701100040H2 SOYMON028 g3021337 BLASTN 602 1e−53 85 1881 16 700895328H1 SOYMON027 g3021337 BLASTN 742 1e−53 83 1882 16 701141083H1 SOYMON038 g3021337 BLASTN 751 1e−53 85 1883 16 700829878H1 SOYMON019 g3021337 BLASTN 417 1e−52 86 1884 16 700671825H1 SOYMON006 g3021337 BLASTN 431 1e−52 79 1885 16 700755240H1 SOYMON014 g3021337 BLASTN 731 1e−52 88 1886 16 701011659H1 SOYMON019 g3021337 BLASTN 734 1e−52 86 1887 16 701011547H1 SOYMON019 g3021337 BLASTN 381 1e−51 84 1888 16 700835614H1 SOYMON019 g3021337 BLASTN 437 1e−51 80 1889 16 700671849H1 SOYMON006 g3021337 BLASTN 471 1e−51 87 1890 16 700734822H1 SOYMON010 g3021337 BLASTN 486 1e−51 79 1891 16 700830223H1 SOYMON019 g3021337 BLASTN 622 1e−51 84 1892 16 700659970H1 SOYMON004 g3021337 BLASTN 722 1e−51 82 1893 16 701101779H1 SOYMON028 g3021337 BLASTN 728 1e−51 86 1894 16 700852553H1 SOYMON023 g3021337 BLASTN 490 1e−50 88 1895 16 700853857H1 SOYMON023 g3021337 BLASTN 711 1e−50 88 1896 16 700980358H1 SOYMON009 g3021337 BLASTN 712 1e−50 85 1897 16 700672182H1 SOYMON006 g3021337 BLASTN 714 1e−50 89 1898 16 700748455H1 SOYMON013 g3021337 BLASTN 396 1e−49 85 1899 16 700657257H1 SOYMON004 g3021337 BLASTN 694 1e−49 75 1900 16 700729301H1 SOYMON009 g3021337 BLASTN 702 1e−49 80 1901 16 700726175H1 SOYMON009 g3021337 BLASTN 704 1e−49 80 1902 16 700966844H1 SOYMON028 g3021337 BLASTN 414 1e−47 81 1903 16 700960965H1 SOYMON022 g3021337 BLASTN 452 1e−47 85 1904 16 700678326H1 SOYMON007 g3021337 BLASTN 480 1e−47 83 1905 16 700751042H1 SOYMON014 g3021337 BLASTN 675 1e−47 87 1906 16 700830863H1 SOYMON019 g3021337 BLASTN 343 1e−46 84 1907 16 701213640H1 SOYMON035 g3021337 BLASTN 667 1e−46 87 1908 16 700870215H1 SOYMON016 g3021337 BLASTN 667 1e−46 80 1909 16 700658278H1 SOYMON004 g3021337 BLASTN 425 1e−44 87 1910 16 700942532H1 SOYMON024 g3021337 BLASTN 583 1e−44 83 1911 16 700986276H1 SOYMON009 g3021337 BLASTN 630 1e−43 81 1912 16 700870216H1 SOYMON016 g3021337 BLASTN 457 1e−42 82 1913 16 700899828H1 SOYMON027 g3021337 BLASTN 464 1e−42 83 1914 16 700678816H1 SOYMON007 g3021337 BLASTN 618 1e−42 86 1915 16 700666809H1 SOYMON005 g3021337 BLASTN 621 1e−42 82 1916 16 701098073H1 SOYMON028 g3021337 BLASTN 285 1e−41 83 1917 16 700669492H1 SOYMON006 g3021337 BLASTN 504 1e−39 83 1918 16 700975340H1 SOYMON009 g3021337 BLASTN 574 1e−39 81 1919 16 700753528H1 SOYMON014 g3021337 BLASTN 576 1e−39 81 1920 16 700665923H1 SOYMON005 g3021337 BLASTN 373 1e−35 84 1921 16 701038320H1 SOYMON029 g3021337 BLASTN 518 1e−34 84 1922 16 700755605H1 SOYMON014 g3021337 BLASTN 431 1e−33 81 1923 16 700890349H1 SOYMON024 g3021337 BLASTN 511 1e−33 88 1924 16 700669817H1 SOYMON006 g3021337 BLASTN 363 1e−31 87 1925 16 701097640H1 SOYMON028 g3021337 BLASTN 476 1e−30 67 1926 16 700562959H1 SOYMON002 g3021337 BLASTN 482 1e−30 81 1927 16 700852454H1 SOYMON023 g3021337 BLASTN 446 1e−28 77 1928 16 701121443H1 SOYMON037 g3021337 BLASTN 418 1e−24 84 1929 16 701118247H1 SOYMON037 g3021337 BLASTN 280 1e−18 85 1930 16 700665401H1 SOYMON005 g927505 BLASTX 172 1e−16 94 1931 16 700750038H1 SOYMON013 g3021338 BLASTX 162 1e−15 84 1932 16 700665414H1 SOYMON005 g3021337 BLASTN 273 1e−13 88 1933 16 700889072H1 SOYMON024 g3021338 BLASTX 136 1e−11 83 1934 16 700727964H1 SOYMON009 g927505 BLASTX 137 1e−11 86 1935 16 700680648H1 SOYMON008 g3021337 BLASTN 226 1e−10 73 1936 16 701044547H1 SOYMON032 g927505 BLASTX 91 1e−9 76 1937 16 700649174H1 SOYMON003 g3021338 BLASTX 126 1e−9 83 1938 16531 701120682H1 SOYMON037 g3021337 BLASTN 716 1e−50 77 1939 1701 700993909H1 SOYMON011 g22633 BLASTX 112 1e−31 78 1940 1701 700955490H1 SOYMON022 g22633 BLASTX 176 1e−25 70 1941 1701 700682081H1 SOYMON008 g22633 BLASTX 147 1e−20 68 1942 1701 700988843H1 SOYMON011 g22633 BLASTX 90 1e−14 67 1943 1701 700740531H1 SOYMON012 g22633 BLASTX 92 1e−12 64 1944 1701 700790059H2 SOYMON011 g22633 BLASTX 92 1e−12 67 1945 1701 700872670H1 SOYMON018 g169037 BLASTX 144 1e−12 90 1946 1701 700990591H1 SOYMON011 g22632 BLASTN 199 1e−11 68 1947 1701 700743120H1 SOYMON012 g22633 BLASTX 92 1e−9 68 1948 1701 700994931H1 SOYMON011 g22633 BLASTX 92 1e−8 64 1949 1938 700738074H1 SOYMON012 g927507 BLASTX 134 1e−11 90 1950 239 701126904H1 SOYMON037 g169037 BLASTX 231 1e−24 81 1951 239 700668532H1 SOYMON006 g169037 BLASTX 202 1e−20 83 1952 239 700943660H1 SOYMON024 g169037 BLASTX 180 1e−17 84 1953 239 701009915H2 SOYMON019 g169037 BLASTX 180 1e−17 84 1954 239 701100047H2 SOYMON028 g169037 BLASTX 160 1e−15 84 1955 239 700794458H1 SOYMON017 g22633 BLASTX 131 1e−10 58 1956 239 700738441H1 SOYMON012 g169037 BLASTX 118 1e−8 78 1957 3425 700984050H1 SOYMON009 g3021337 BLASTN 874 1e−64 80 1958 3425 701014509H1 SOYMON019 g3021337 BLASTN 520 1e−60 80 1959 3425 701138819H1 SOYMON038 g3021337 BLASTN 815 1e−59 80 1960 3425 700977309H1 SOYMON009 g3021337 BLASTN 809 1e−58 80 1961 3425 700984876H1 SOYMON009 g3021337 BLASTN 813 1e−58 80 1962 3425 701046151H1 SOYMON032 g3021337 BLASTN 730 1e−52 80 1963 3425 700889668H1 SOYMON024 g3021337 BLASTN 737 1e−52 81 1964 3425 700976571H1 SOYMON009 g3021337 BLASTN 737 1e−52 81 1965 3425 701045371H1 SOYMON032 g3021337 BLASTN 716 1e−50 79 1966 3425 700548283H1 SOYMON002 g3021337 BLASTN 700 1e−49 81 1967 3425 701103461H1 SOYMON028 g3021337 BLASTN 705 1e−49 81 1968 3425 700898446H1 SOYMON027 g3021337 BLASTN 686 1e−48 83 1969 3425 701006432H1 SOYMON019 g3021337 BLASTN 688 1e−48 83 1970 3425 701041476H1 SOYMON029 g3021337 BLASTN 693 1e−48 81 1971 3425 700568335H1 SOYMON002 g3021337 BLASTN 678 1e−47 82 1972 3425 701046312H1 SOYMON032 g3021337 BLASTN 650 1e−45 85 1973 3425 701050171H1 SOYMON032 g3021337 BLASTN 650 1e−45 85 1974 3425 700685063H1 SOYMON008 g3021337 BLASTN 643 1e−44 83 1975 3425 701010250H2 SOYMON019 g3021337 BLASTN 542 1e−36 86 1976 3425 700665454H1 SOYMON005 g3021337 BLASTN 520 1e−34 80 1977 3425 701043888H1 SOYMON032 g3021337 BLASTN 495 1e−32 85 1978 3425 700726806H1 SOYMON009 g3021337 BLASTN 213 1e−23 76 1979 491 700997879H1 SOYMON018 g22632 BLASTN 789 1e−56 77 1980 491 700646208H1 SOYMON012 g22632 BLASTN 733 1e−52 76 1981 491 700559796H1 SOYMON001 g22632 BLASTN 715 1e−50 76 1982 491 700789784H1 SOYMON011 g22632 BLASTN 664 1e−46 76 1983 491 700683122H1 SOYMON008 g22632 BLASTN 485 1e−41 86 1984 491 701105914H1 SOYMON036 g22632 BLASTN 504 1e−41 73 1985 491 700558789H1 SOYMON001 g22632 BLASTN 607 1e−41 74 1986 491 700873051H1 SOYMON018 g22632 BLASTN 608 1e−41 75 1987 491 700684010H1 SOYMON008 g22632 BLASTN 597 1e−40 75 1988 491 700786096H2 SOYMON011 g22632 BLASTN 576 1e−39 75 1989 491 700731865H1 SOYMON010 g22632 BLASTN 582 1e−39 75 1990 491 701108111H1 SOYMON036 g22632 BLASTN 467 1e−38 75 1991 491 700740887H1 SOYMON012 g22632 BLASTN 567 1e−38 74 1992 491 700559579H1 SOYMON001 g22632 BLASTN 572 1e−38 75 1993 491 700996104H1 SOYMON018 g22632 BLASTN 476 1e−37 76 1994 491 700682145H1 SOYMON008 g22632 BLASTN 542 1e−36 74 1995 491 700737263H1 SOYMON010 g22632 BLASTN 526 1e−35 74 1996 491 700547963H1 SOYMON001 g22632 BLASTN 527 1e−35 73 1997 491 700686296H1 SOYMON008 g22632 BLASTN 527 1e−35 73 1998 491 700646072H1 SOYMON011 g22632 BLASTN 537 1e−35 74 1999 491 701106662H1 SOYMON036 g22632 BLASTN 514 1e−34 74 2000 491 700684335H1 SOYMON008 g22632 BLASTN 516 1e−34 74 2001 491 701000609H1 SOYMON018 g22632 BLASTN 520 1e−34 74 2002 491 700685658H1 SOYMON008 g22632 BLASTN 520 1e−34 74 2003 491 700875532H1 SOYMON018 g22632 BLASTN 521 1e−34 73 2004 491 700730264H1 SOYMON009 g22632 BLASTN 502 1e−33 74 2005 491 700872948H1 SOYMON018 g22632 BLASTN 502 1e−33 74 2006 491 700685813H1 SOYMON008 g22632 BLASTN 502 1e−33 74 2007 491 701104554H1 SOYMON036 g22632 BLASTN 503 1e−33 74 2008 491 700960601H1 SOYMON022 g22632 BLASTN 503 1e−33 74 2009 491 700876633H1 SOYMON018 g22632 BLASTN 503 1e−33 74 2010 491 700739662H1 SOYMON012 g22632 BLASTN 504 1e−33 72 2011 491 700685904H1 SOYMON008 g22632 BLASTN 505 1e−33 72 2012 491 700995183H1 SOYMON011 g22632 BLASTN 513 1e−33 73 2013 491 700901996H1 SOYMON027 g22632 BLASTN 513 1e−33 74 2014 491 700727070H1 SOYMON009 g22632 BLASTN 490 1e−32 72 2015 491 700685790H1 SOYMON008 g22632 BLASTN 492 1e−32 74 2016 491 700998652H1 SOYMON018 g22632 BLASTN 494 1e−32 72 2017 491 700740465H1 SOYMON012 g22632 BLASTN 482 1e−31 74 2018 491 700682621H2 SOYMON008 g22632 BLASTN 484 1e−31 74 2019 491 700874316H1 SOYMON018 g22632 BLASTN 466 1e−30 73 2020 491 700686477H1 SOYMON008 g22632 BLASTN 473 1e−30 73 2021 491 700739979H1 SOYMON012 g22632 BLASTN 476 1e−30 74 2022 491 700739416H1 SOYMON012 g22632 BLASTN 476 1e−30 74 2023 491 700685976H1 SOYMON008 g22632 BLASTN 476 1e−30 74 2024 491 700739629H1 SOYMON012 g22632 BLASTN 486 1e−30 70 2025 491 700989163H1 SOYMON011 g22632 BLASTN 468 1e−29 72 2026 491 701000555H1 SOYMON018 g22632 BLASTN 477 1e−29 72 2027 491 700872702H1 SOYMON018 g22632 BLASTN 436 1e−28 72 2028 491 701000781H1 SOYMON018 g22632 BLASTN 460 1e−28 73 2029 491 700682760H1 SOYMON008 g22632 BLASTN 463 1e−28 72 2030 491 700740390H1 SOYMON012 g22632 BLASTN 440 1e−27 73 2031 491 700685346H1 SOYMON008 g22632 BLASTN 451 1e−27 72 2032 491 700557272H1 SOYMON001 g22632 BLASTN 250 1e−26 78 2033 491 700953343H1 SOYMON022 g22632 BLASTN 349 1e−26 74 2034 491 700741960H1 SOYMON012 g22632 BLASTN 430 1e−26 73 2035 491 700680247H2 SOYMON008 g22632 BLASTN 425 1e−25 67 2036 491 700680002H2 SOYMON008 g22632 BLASTN 241 1e−24 72 2037 491 700684827H1 SOYMON008 g22632 BLASTN 379 1e−24 74 2038 491 700956353H1 SOYMON022 g22632 BLASTN 410 1e−24 72 2039 491 700787513H1 SOYMON011 g22632 BLASTN 235 1e−22 72 2040 491 700725070H1 SOYMON009 g22632 BLASTN 241 1e−22 71 2041 491 700741111H1 SOYMON012 g22632 BLASTN 304 1e−22 73 2042 491 700985308H1 SOYMON009 g22632 BLASTN 241 1e−21 80 2043 491 700738230H1 SOYMON012 g22632 BLASTN 241 1e−21 72 2044 491 700991396H1 SOYMON011 g22632 BLASTN 350 1e−21 72 2045 491 700741276H1 SOYMON012 g22632 BLASTN 379 1e−21 71 2046 491 700740223H1 SOYMON012 g22632 BLASTN 241 1e−20 72 2047 491 700738808H1 SOYMON012 g22632 BLASTN 241 1e−20 72 2048 491 700997995H1 SOYMON018 g22632 BLASTN 241 1e−19 81 2049 491 700875139H1 SOYMON018 g22632 BLASTN 241 1e−19 71 2050 491 700989713H1 SOYMON011 g22632 BLASTN 241 1e−19 73 2051 491 700958366H1 SOYMON022 g22632 BLASTN 241 1e−18 71 2052 491 700683887H1 SOYMON008 g22632 BLASTN 344 1e−18 70 2053 491 700740788H1 SOYMON012 g22632 BLASTN 339 1e−17 70 2054 491 700743058H1 SOYMON012 g22632 BLASTN 205 1e−16 81 2055 491 700996423H1 SOYMON018 g22632 BLASTN 234 1e−16 80 2056 491 700686075H1 SOYMON008 g22632 BLASTN 241 1e−16 71 2057 491 700738811H1 SOYMON012 g22632 BLASTN 193 1e−15 72 2058 491 700998312H1 SOYMON018 g22632 BLASTN 234 1e−15 73 2059 491 700681825H1 SOYMON008 g22632 BLASTN 241 1e−15 81 2060 491 701109105H1 SOYMON036 g22632 BLASTN 290 1e−14 69 2061 491 701203741H2 SOYMON035 g22632 BLASTN 230 1e−13 78 2062 491 700740785H1 SOYMON012 g22632 BLASTN 287 1e−13 68 2063 491 700738486H1 SOYMON012 g22632 BLASTN 295 1e−13 64 2064 491 700739078H1 SOYMON012 g22632 BLASTN 178 1e−12 73 2065 491 701002287H1 SOYMON018 g22632 BLASTN 255 1e−12 74 2066 491 700742470H1 SOYMON012 g22632 BLASTN 278 1e−12 69 2067 491 700743421H1 SOYMON012 g22632 BLASTN 261 1e−11 71 2068 491 700744039H1 SOYMON012 g22632 BLASTN 265 1e−11 69 2069 491 700789444H2 SOYMON011 g22632 BLASTN 158 1e−10 87 2070 491 700741074H1 SOYMON012 g22632 BLASTN 178 1e−10 77 2071 491 700998877H1 SOYMON018 g22632 BLASTN 235 1e−10 72 2072 491 700740005H1 SOYMON012 g22633 BLASTX 75 1e−9 64 2073 491 700872703H1 SOYMON018 g169037 BLASTX 116 1e−9 83 2074 491 700743301H1 SOYMON012 g22632 BLASTN 241 1e−9 76 2075 491 700875039H1 SOYMON018 g22632 BLASTN 241 1e−9 72 2076 491 700742515H1 SOYMON012 g22632 BLASTN 241 1e−9 76 2077 491 700990557H1 SOYMON011 g22632 BLASTN 241 1e−9 76 2078 491 700743995H1 SOYMON012 g22632 BLASTN 241 1e−9 76 2079 491 700743495H1 SOYMON012 g22632 BLASTN 241 1e−9 76 2080 491 701001909H1 SOYMON018 g22632 BLASTN 241 1e−9 76 2081 491 701001445H1 SOYMON018 g169037 BLASTX 115 1e−8 92 2082 491 700554881H1 SOYMON001 g169037 BLASTX 116 1e−8 94 2083 491 700954194H1 SOYMON022 g169037 BLASTX 116 1e−8 94 2084 491 700996869H1 SOYMON018 g22632 BLASTN 230 1e−8 76 2085 491 700897820H1 SOYMON027 g22632 BLASTN 234 1e−8 74 2086 491 700742574H1 SOYMON012 g22632 BLASTN 234 1e−8 74 2087 491 700684738H1 SOYMON008 g22632 BLASTN 235 1e−8 75 2088 7368 700739343H1 SOYMON012 g927507 BLASTX 164 1e−15 88 2089 −GM32379 LIB3051-015- LIB3051 g3021337 BLASTN 260 1e−28 77 Q1-E1-B12 2090 −GM8265 LIB3039-048- LIB3039 g3021337 BLASTN 481 1e−29 65 Q1-E1-F11 2091 16 LIB3027-010- LIB3027 g3021337 BLASTN 1393 1e−107 82 Q1-B1-B7 2092 16 LIB3039-049- LIB3039 g3021337 BLASTN 1297 1e−99 83 Q1-E1-B8 2093 16 LIB3051-061- LIB3051 g3021337 BLASTN 1303 1e−99 84 Q1-K1-E11 2094 16 LIB3056-009- LIB3056 g3021337 BLASTN 1126 1e−96 84 Q1-N1-A10 2095 16 LIB3051-025- LIB3051 g3021337 BLASTN 1262 1e−96 83 Q1-K1-E11 2096 16 LIB3056-014- LIB3056 g3021337 BLASTN 1077 1e−94 81 Q1-N1-E1 2097 16 LIB3055-005- LIB3055 g3021337 BLASTN 1227 1e−93 84 Q1-N1-A8 2098 16 LIB3040-045- LIB3040 g3021337 BLASTN 1211 1e−92 83 Q1-E1-A4 2099 16 LIB3028-010- LIB3028 g3021337 BLASTN 1215 1e−92 83 Q1-B1-G9 2100 16 LIB3056-010- LIB3056 g3021337 BLASTN 1217 1e−92 84 Q1-N1-G8 2101 16 LIB3039-029- LIB3039 g3021337 BLASTN 1128 1e−85 85 Q1-E1-A6 2102 16 LIB3051-014- LIB3051 g3021337 BLASTN 716 1e−80 83 Q1-E1-D2 2103 16 LIB3030-010- LIB3030 g3021337 BLASTN 1052 1e−78 83 Q1-B1-D7 2104 16 LIB3051-094- LIB3051 g3021337 BLASTN 778 1e−74 83 Q1-K1-A9 2105 16 LIB3028-030- LIB3028 g3021337 BLASTN 953 1e−70 85 Q1-B1-C9 2106 16 LIB3052-004- LIB3052 g3021337 BLASTN 868 1e−63 82 Q1-N1-D8 2107 16 LIB3065-014- LIB3065 g3021337 BLASTN 540 1e−61 79 Q1-N1-A3 2108 16 LIB3050-019- LIB3050 g168420 BLASTX 223 1e−40 63 Q1-K1-H1 2109 16 LIB3051-062- LIB3051 g3021337 BLASTN 541 1e−38 79 Q1-K1-B5 2110 3425 LIB3051-067- LIB3051 g3021337 BLASTN 1082 1e−81 78 Q1-K1-E7 2111 3425 LIB3050-006- LIB3050 g3021337 BLASTN 752 1e−57 75 Q1-E1-G7 2112 491 LIB3028-011- LIB3028 g22632 BLASTN 911 1e−67 75 Q1-B1-B9 2113 491 LIB3028-011- LIB3028 g22632 BLASTN 886 1e−65 77 Q1-B1-F2 SOYBEAN FRUCTOSE-1,6-BISPHOSPHATASE 2114 −700685384 700685384H1 SOYMON008 g21244 BLASTN 597 1e−49 80 2115 −700737915 700737915H1 SOYMON012 g515746 BLASTN 1316 1e−100 97 2116 −700741457 700741457H1 SOYMON012 g3041774 BLASTN 692 1e−58 80 2117 −700874831 700874831H1 SOYMON018 g515746 BLASTN 1295 1e−99 100 2118 −700996155 700996155H1 SOYMON018 g3041774 BLASTN 651 1e−45 83 2119 −700996632 700996632H1 SOYMON018 g515746 BLASTN 507 1e−51 90 2120 −700998027 700998027H1 SOYMON018 g515746 BLASTN 636 1e−65 94 2121 −701209548 701209548H1 SOYMON035 g3041774 BLASTN 642 1e−44 83 2122 10129 700870828H1 SOYMON018 g21244 BLASTN 827 1e−60 79 2123 10129 700741669H1 SOYMON012 g21244 BLASTN 657 1e−53 80 2124 10348 700555754H1 SOYMON001 g21244 BLASTN 466 1e−29 77 2125 10348 700991527H1 SOYMON011 g440591 BLASTX 169 1e−16 88 2126 13716 700898719H1 SOYMON027 g515746 BLASTN 1186 1e−90 97 2127 13716 700993540H1 SOYMON011 g515746 BLASTN 1179 1e−89 98 2128 13716 700909657H1 SOYMON022 g515746 BLASTN 568 1e−57 86 2129 1894 700555054H1 SOYMON001 g515746 BLASTN 1320 1e−101 100 2130 1894 700685264H1 SOYMON008 g515746 BLASTN 1323 1e−101 99 2131 1894 700558854H1 SOYMON001 g515746 BLASTN 695 1e−98 100 2132 1894 700554755H1 SOYMON001 g515746 BLASTN 767 1e−98 99 2133 1894 701000504H1 SOYMON018 g515746 BLASTN 626 1e−95 98 2134 1894 700738115H1 SOYMON012 g515746 BLASTN 1230 1e−93 100 2135 1894 700992933H1 SOYMON011 g515746 BLASTN 1074 1e−91 98 2136 1894 701107444H1 SOYMON036 g515746 BLASTN 1201 1e−91 99 2137 1894 700852823H1 SOYMON023 g515746 BLASTN 1041 1e−90 98 2138 1894 700733478H1 SOYMON010 g515746 BLASTN 1150 1e−90 97 2139 1894 701105185H1 SOYMON036 g515746 BLASTN 641 1e−87 89 2140 1894 700737830H1 SOYMON012 g515746 BLASTN 1060 1e−87 100 2141 1894 700685110H1 SOYMON008 g515746 BLASTN 597 1e−86 90 2142 1894 700968307H1 SOYMON036 g515746 BLASTN 1113 1e−84 97 2143 1894 700653014H1 SOYMON003 g515746 BLASTN 587 1e−82 90 2144 1894 700555504H1 SOYMON001 g515746 BLASTN 626 1e−81 88 2145 1894 700751540H1 SOYMON014 g515746 BLASTN 585 1e−77 91 2146 1894 700901976H1 SOYMON027 g515746 BLASTN 505 1e−73 87 2147 1894 700986496H1 SOYMON009 g515746 BLASTN 559 1e−73 90 2148 1894 700751580H1 SOYMON014 g515746 BLASTN 569 1e−72 89 2149 1894 700751532H1 SOYMON014 g515746 BLASTN 571 1e−72 90 2150 1894 700990937H1 SOYMON011 g515746 BLASTN 544 1e−71 88 2151 1894 700740789H1 SOYMON012 g515746 BLASTN 630 1e−69 100 2152 1894 700743994H1 SOYMON012 g515746 BLASTN 945 1e−69 100 2153 1894 700754374H1 SOYMON014 g515746 BLASTN 460 1e−62 91 2154 1894 701001295H1 SOYMON018 g515746 BLASTN 541 1e−62 97 2155 1894 701155952H1 SOYMON031 g515746 BLASTN 568 1e−51 83 2156 1894 700872212H1 SOYMON018 g515746 BLASTN 670 1e−47 100 2157 1894 700682196H1 SOYMON008 g515746 BLASTN 609 1e−41 98 2158 1894 700738779H1 SOYMON012 g515746 BLASTN 252 1e−16 82 2159 26568 700844816H1 SOYMON021 g21244 BLASTN 649 1e−45 78 2160 27512 701128049H1 SOYMON037 g440591 BLASTX 185 1e−18 87 2161 7128 700649846H1 SOYMON003 g440591 BLASTX 125 1e−15 81 2162 10348 LIB3030-010- LIB3030 g21244 BLASTN 476 1e−28 76 Q1-B1-C7 FRUCTOSE-6-PHOSPHATE,2-KINASE 2163 −700730441 700730441H1 SOYMON009 g3309583 BLASTX 179 1e−17 82 2164 −700953509 700953509H1 SOYMON022 g3170229 BLASTN 674 1e−47 75 2165 −700955121 700955121H1 SOYMON022 g3309582 BLASTN 303 1e−14 68 2166 −GM28972 LIB3050-012- LIB3050 g3170229 BLASTN 1073 1e−80 80 Q1-E1-E9 SOYBEAN PHOSPHOGLUCOISOMERASE 2167 −700568558 700568558H1 SOYMON002 g1369950 BLASTX 165 1e−15 80 2168 −700845275 700845275H1 SOYMON021 g1100771 BLASTX 124 1e−10 53 2169 −700960755 700960755H1 SOYMON022 g1100771 BLASTX 153 1e−14 52 2170 18663 700838363H1 SOYMON020 g1100771 BLASTX 215 1e−22 63 2171 18663 700838355H1 SOYMON020 g1100771 BLASTX 155 1e−14 81 2172 19355 700897450H1 SOYMON027 g1100771 BLASTX 273 1e−31 74 2173 19355 700744258H1 SOYMON013 g1100771 BLASTX 207 1e−29 69 2174 19355 701153832H1 SOYMON031 g1100771 BLASTX 226 1e−23 58 2175 20088 700856114H1 SOYMON023 g1100771 BLASTX 176 1e−33 75 2176 20088 700670380H1 SOYMON006 g1100771 BLASTX 207 1e−33 71 2177 20088 700788785H2 SOYMON011 g1100771 BLASTX 120 1e−32 74 2178 20088 700847659H1 SOYMON021 g1100771 BLASTX 192 1e−31 84 2179 20088 701136417H1 SOYMON038 g1100771 BLASTX 169 1e−27 66 2180 31255 701207622H1 SOYMON035 g1100771 BLASTX 168 1e−29 61 2181 20088 LIB3051-014- LIB3051 g1100771 BLASTX 400 1e−68 73 Q1-E1-G3 2182 31255 LIB3056-008- LIB3056 g1100771 BLASTX 188 1e−52 62 Q1-N1-G8 SOYBEAN VACUOLAR H+-TRANSLOCATING-PYROPHOSPHATASE 2183 −700660662 700660662H1 SOYMON004 g16347 BLASTN 540 1e−36 79 2184 −700793860 700793860H1 SOYMON017 g2706449 BLASTN 808 1e−58 78 2185 −700837007 700837007H1 SOYMON020 g16347 BLASTN 776 1e−55 78 2186 −700890647 700890647H1 SOYMON024 g790474 BLASTN 826 1e−60 81 2187 −700942978 700942978H1 SOYMON024 g790478 BLASTN 605 1e−63 82 2188 −700944280 700944280H1 SOYMON024 g790479 BLASTX 119 1e−10 76 2189 −700974544 700974544H1 SOYMON005 g1103711 BLASTN 854 1e−62 83 2190 −700984449 700984449H1 SOYMON009 g1103711 BLASTN 287 1e−12 71 2191 −700989248 700989248H1 SOYMON011 g534915 BLASTN 276 1e−14 67 2192 −701102931 701102931H1 SOYMON028 g2706449 BLASTN 438 1e−46 76 2193 −701106870 701106870H1 SOYMON036 g790478 BLASTN 623 1e−47 75 2194 −701122796 701122796H1 SOYMON037 g2258074 BLASTX 71 1e−15 73 2195 −701132123 701132123H1 SOYMON038 g790478 BLASTN 627 1e−43 81 2196 −701136557 701136557H1 SOYMON038 g16347 BLASTN 376 1e−33 77 2197 14021 700973215H1 SOYMON005 g2668745 BLASTN 435 1e−39 80 2198 14021 701109310H1 SOYMON036 g2668745 BLASTN 281 1e−25 83 2199 16 700891764H1 SOYMON024 g790479 BLASTX 172 1e−16 68 2200 19232 701061126H1 SOYMON033 g790474 BLASTN 935 1e−69 81 2201 19232 700962864H1 SOYMON022 g790474 BLASTN 874 1e−64 82 2202 20872 700754883H1 SOYMON014 g790478 BLASTN 824 1e−59 81 2203 20872 700971147H1 SOYMON005 g1103711 BLASTN 564 1e−54 79 2204 2813 700797861H1 SOYMON017 g16347 BLASTN 731 1e−52 79 2205 2813 700944850H1 SOYMON024 g2570500 BLASTN 738 1e−52 82 2206 2813 701056207H1 SOYMON032 g2570500 BLASTN 556 1e−46 80 2207 2813 700605115H2 SOYMON003 g2570500 BLASTN 478 1e−42 80 2208 2813 700897063H1 SOYMON027 g2570500 BLASTN 596 1e−40 80 2209 2813 700561829H1 SOYMON002 g2570500 BLASTN 570 1e−38 80 2210 2813 701204883H1 SOYMON035 g2668745 BLASTN 545 1e−36 77 2211 2813 700754984H1 SOYMON014 g2570500 BLASTN 527 1e−35 75 2212 2813 700854552H1 SOYMON023 g2570500 BLASTN 536 1e−35 79 2213 2813 700873337H1 SOYMON018 g2570500 BLASTN 505 1e−33 75 2214 2813 700873349H1 SOYMON018 g2570500 BLASTN 506 1e−33 75 2215 2813 700952403H1 SOYMON022 g2668745 BLASTN 499 1e−32 76 2216 2813 700846561H1 SOYMON021 g2570500 BLASTN 488 1e−31 75 2217 2813 700953987H1 SOYMON022 g2570500 BLASTN 461 1e−29 75 2218 2813 700568667H1 SOYMON002 g2570500 BLASTN 296 1e−24 79 2219 2813 700895231H1 SOYMON024 g2258074 BLASTX 207 1e−22 80 2220 2813 701101791H1 SOYMON028 g2668746 BLASTX 147 1e−13 77 2221 8040 701121224H1 SOYMON037 g534915 BLASTN 298 1e−14 77 2222 8040 700743066H1 SOYMON012 g2668746 BLASTX 140 1e−12 80 2223 8531 701005139H1 SOYMON019 g2258073 BLASTN 871 1e−63 79 2224 8531 701008308H1 SOYMON019 g534915 BLASTN 789 1e−57 76 2225 8531 700559054H1 SOYMON001 g2570500 BLASTN 790 1e−57 77 2226 8531 700942540H1 SOYMON024 g2706449 BLASTN 755 1e−54 80 2227 8531 700790983H1 SOYMON011 g2258073 BLASTN 431 1e−52 77 2228 8531 701007949H1 SOYMON019 g2570500 BLASTN 404 1e−41 70 2229 8531 701123827H1 SOYMON037 g534915 BLASTN 436 1e−26 75 2230 8531 701013616H1 SOYMON019 g534915 BLASTN 431 1e−25 78 2231 8531 700565624H1 SOYMON002 g2570501 BLASTX 169 1e−16 85 2232 8531 701121092H1 SOYMON037 g2570501 BLASTX 110 1e−15 60 2233 16 LIB3040-003- LIB3040 g633598 BLASTN 523 1e−51 74 Q1-E1-F6 2234 16 LIB3051-114- LIB3051 g790478 BLASTN 457 1e−48 79 Q1-K1-G5 2235 16 LIB3039-020- LIB3039 g790478 BLASTN 338 1e−30 74 Q1-E1-A2 2236 2813 LIB3028-026- LIB3028 g2570500 BLASTN 1029 1e−77 80 Q1-B1-B7 2237 8040 LIB3049-045- LIB3049 g2706449 BLASTN 752 1e−52 72 Q1-E1 -C3 2238 8040 LIB3049-005- LIB3049 g2570501 BLASTX 154 1e−32 61 Q1-E1-A7 2239 8531 LIB3050-013- LIB3050 g2570500 BLASTN 748 1e−53 72 Q1-E1 -G8 2240 8531 LIB3073-025- LIB3073 g534915 BLASTN 711 1e−49 78 Q1-K1-D6 2241 8531 LIB3050-012- LIB3050 g2258074 BLASTX 93 1e−31 74 Q1-E1-D1 SOYBEAN PYROPHOSPHATE-DEPENDENT FRUCTOSE-6-PHOSPHATE PHOSPHOTRANSFERASE 2242 7899 701008645H1 SOYMON019 g169538 BLASTX 160 1e−15 83 INVERTASES 2243 −700653543 700653543H1 SOYMON003 g1160487 BLASTN 541 1e−55 84 2244 −700992760 700992760H1 SOYMON011 g550319 BLASTX 117 1e−12 49 2245 −701005703 701005703H1 SOYMON019 g861157 BLASTX 213 1e−22 46 2246 −701047324 701047324H1 SOYMON032 g1160487 BLASTN 647 1e−45 81 2247 −701130328 701130328H1 SOYMON037 g167551 BLASTX 215 1e−22 61 2248 20460 700658149H1 SOYMON004 g861157 BLASTX 198 1e−20 72 2249 20460 701041452H1 SOYMON029 g402740 BLASTX 105 1e−13 76 2250 −GM31611 LIB3051-002- LIB3051 g1160487 BLASTN 1033 1e−77 77 Q1-E1-B9 2251 −GM34282 LIB3051-025- LIB3051 g1160487 BLASTN 1069 1e−80 79 Q1-K1-C4 2252 −GM34976 LIB3051-031- LIB3051 g1160487 BLASTN 769 1e−66 80 Q1-K1-A9 2253 31949 LIB3051-093- LIB3051 g1160487 BLASTN 948 1e−92 77 Q1-K1-B1 2254 31949 LIB3051-054- LIB3051 g1160487 BLASTN 903 1e−90 82 Q1-K2-D11 SOYBEAN SUCROSE SYNTHASE 2255 −700565776 700565776H1 SOYMON002 g3169544 BLASTX 89 1e−8 64 2256 −700606005 700606005H2 SOYMON007 g2570066 BLASTN 1069 1e−80 89 2257 −700664186 700664186H1 SOYMON005 g2606080 BLASTN 426 1e−62 91 2258 −700668119 700668119H1 SOYMON006 g2570066 BLASTN 279 1e−14 83 2259 −700668348 700668348H1 SOYMON006 g2570066 BLASTN 693 1e−48 88 2260 −700671225 700671225H1 SOYMON006 g16525 BLASTN 617 1e−42 72 2261 −700673918 700673918H1 SOYMON007 g218332 BLASTN 152 1e−9 92 2262 −700726266 700726266H1 SOYMON009 g2606080 BLASTN 237 1e−21 79 2263 −700747171 700747171H1 SOYMON013 g2606080 BLASTN 735 1e−52 89 2264 −700747359 700747359H1 SOYMON013 g218332 BLASTN 447 1e−28 78 2265 −700787443 700787443H2 SOYMON011 g22485 BLASTN 1171 1e−95 98 2266 −700796035 700796035H1 SOYMON017 g2570066 BLASTN 1039 1e−77 90 2267 −700832792 700832792H1 SOYMON019 g2606080 BLASTN 444 1e−31 88 2268 −700836673 700836673H1 SOYMON020 g2570066 BLASTN 843 1e−61 85 2269 −700841855 700841855H1 SOYMON020 g2570066 BLASTN 425 1e−35 84 2270 −700851758 700851758H1 SOYMON023 g2570066 BLASTN 211 1e−15 91 2271 −700851991 700851991H1 SOYMON023 g2570066 BLASTN 768 1e−55 81 2272 −700852943 700852943H1 SOYMON023 g2606080 BLASTN 250 1e−13 85 2273 −700853396 700853396H1 SOYMON023 g2570067 BLASTX 145 1e−13 65 2274 −700872206 700872206H1 SOYMON018 g1488570 BLASTX 235 1e−25 64 2275 −700876641 700876641H1 SOYMON018 g2606080 BLASTN 410 1e−53 88 2276 −700890526 700890526H1 SOYMON024 g2606080 BLASTN 652 1e−60 83 2277 −700893784 700893784H1 SOYMON024 g3169543 BLASTN 217 1e−11 82 2278 −700909222 700909222H1 SOYMON022 g2570066 BLASTN 440 1e−44 72 2279 −700944438 700944438H1 SOYMON024 g3169543 BLASTN 669 1e−46 73 2280 −700945733 700945733H1 SOYMON024 g1488569 BLASTN 504 1e−33 66 2281 −700969926 700969926H1 SOYMON005 g2570066 BLASTN 674 1e−47 72 2282 −701001986 701001986H1 SOYMON018 g1146237 BLASTX 106 1e−9 45 2283 −701005687 701005687H1 SOYMON019 g2606080 BLASTN 591 1e−40 85 2284 −701012195 701012195H1 SOYMON019 g2606080 BLASTN 418 1e−46 77 2285 −701046403 701046403H1 SOYMON032 g2606080 BLASTN 574 1e−38 76 2286 −701058966 701058966H1 SOYMON033 g218332 BLASTN 529 1e−56 84 2287 −701150574 701150574H1 SOYMON031 g1041247 BLASTX 155 1e−14 74 2288 −701205210 701205210H1 SOYMON035 g218332 BLASTN 981 1e−72 85 2289 10445 700605276H2 SOYMON003 g2606080 BLASTN 860 1e−65 84 2290 10445 700832417H1 SOYMON019 g2606080 BLASTN 876 1e−64 82 2291 10445 700833214H1 SOYMON019 g2606080 BLASTN 740 1e−58 83 2292 10445 700832409H1 SOYMON019 g2606080 BLASTN 800 1e−57 84 2293 10445 701007169H1 SOYMON019 g2606080 BLASTN 691 1e−55 81 2294 10445 701005913H1 SOYMON019 g2606080 BLASTN 680 1e−52 83 2295 10445 701204549H2 SOYMON035 g2606080 BLASTN 732 1e−52 83 2296 10445 701208347H1 SOYMON035 g2606080 BLASTN 656 1e−49 83 2297 10445 700958980H1 SOYMON022 g2606080 BLASTN 670 1e−49 83 2298 10445 700988126H1 SOYMON009 g2606080 BLASTN 324 1e−47 78 2299 10445 700830464H1 SOYMON019 g2606080 BLASTN 347 1e−47 79 2300 10445 700763911H1 SOYMON019 g3169543 BLASTN 517 1e−47 75 2301 10445 700891996H1 SOYMON024 g2606080 BLASTN 667 1e−46 88 2302 10445 700725104H1 SOYMON009 g2606080 BLASTN 577 1e−45 81 2303 10445 701124001H1 SOYMON037 g2606080 BLASTN 648 1e−45 86 2304 10445 700833919H1 SOYMON019 g2606080 BLASTN 496 1e−41 79 2305 10445 701006692H1 SOYMON019 g2606080 BLASTN 536 1e−41 86 2306 10445 700905349H1 SOYMON022 g2606080 BLASTN 585 1e−39 75 2307 10445 701204596H2 SOYMON035 g2606080 BLASTN 521 1e−38 79 2308 10445 700958885H1 SOYMON022 g2606080 BLASTN 351 1e−36 81 2309 10445 701208390H1 SOYMON035 g2606080 BLASTN 259 1e−29 86 2310 10445 701003131H1 SOYMON019 g2606080 BLASTN 442 1e−26 76 2311 10445 701207712H1 SOYMON035 g2606080 BLASTN 260 1e−17 78 2312 10445 701215107H1 SOYMON035 g2606080 BLASTN 260 1e−14 88 2313 10445 700852649H1 SOYMON023 g2606080 BLASTN 254 1e−13 74 2314 11259 701063407H1 SOYMON033 g2570066 BLASTN 1100 1e−82 87 2315 11259 700674761H1 SOYMON007 g2570066 BLASTN 739 1e−71 86 2316 11259 700839148H1 SOYMON020 g2570066 BLASTN 919 1e−67 87 2317 11259 700674815H1 SOYMON007 g2570066 BLASTN 904 1e−66 87 2318 12890 701103318H1 SOYMON028 g2570066 BLASTN 1005 1e−74 86 2319 12890 700855911H1 SOYMON023 g2570066 BLASTN 569 1e−69 86 2320 12890 700850874H1 SOYMON023 g2570066 BLASTN 937 1e−69 90 2321 12890 700837552H1 SOYMON020 g2570066 BLASTN 888 1e−65 89 2322 14264 700677058H1 SOYMON007 g2606080 BLASTN 578 1e−39 99 2323 14264 700679301H1 SOYMON007 g2606080 BLASTN 325 1e−18 90 2324 14740 701214452H1 SOYMON035 g2570066 BLASTN 1072 1e−80 89 2325 14740 701044972H1 SOYMON032 g2570066 BLASTN 537 1e−43 87 2326 14740 701040560H1 SOYMON029 g2570066 BLASTN 302 1e−24 75 2327 14740 700793901H1 SOYMON017 g2570066 BLASTN 231 1e−14 84 2328 15394 701136903H1 SOYMON038 g2606080 BLASTN 936 1e−69 81 2329 15394 701004431H1 SOYMON019 g218332 BLASTN 942 1e−69 80 2330 15394 701006153H1 SOYMON019 g218332 BLASTN 920 1e−67 83 2331 15394 701138281H1 SOYMON038 g218332 BLASTN 485 1e−40 82 2332 15394 701209319H1 SOYMON035 g3169543 BLASTN 508 1e−33 81 2333 16344 700746372H1 SOYMON013 g2606080 BLASTN 471 1e−65 85 2334 16344 700945706H1 SOYMON024 g2606080 BLASTN 635 1e−65 84 2335 17781 700960671H1 SOYMON022 g2570066 BLASTN 966 1e−71 88 2336 17781 700838540H1 SOYMON020 g2570066 BLASTN 532 1e−62 83 2337 20151 700847184H1 SOYMON021 g2570066 BLASTN 762 1e−72 90 2338 20151 700831558H1 SOYMON019 g2570066 BLASTN 980 1e−72 89 2339 22196 701046171H1 SOYMON032 g2606080 BLASTN 1321 1e−101 99 2340 22196 701207390H1 SOYMON035 g2606080 BLASTN 1258 1e−95 98 2341 25275 701013025H1 SOYMON019 g2606080 BLASTN 1353 1e−103 98 2342 25275 700561738H1 SOYMON002 g2606080 BLASTN 953 1e−84 91 2343 25380 700667735H1 SOYMON006 g2570066 BLASTN 959 1e−71 87 2344 25380 701047629H1 SOYMON032 g2570066 BLASTN 774 1e−55 89 2345 26818 701047072H1 SOYMON032 g2606080 BLASTN 830 1e−60 87 2346 26818 700737511H1 SOYMON010 g3169543 BLASTN 607 1e−57 83 2347 31182 701098655H1 SOYMON028 g2570066 BLASTN 951 1e−70 85 2348 318 701052316H1 SOYMON032 g2606080 BLASTN 1555 1e−120 100 2349 318 701053115H1 SOYMON032 g2606080 BLASTN 1281 1e−111 96 2350 318 700983049H1 SOYMON009 g2606080 BLASTN 1438 1e−110 96 2351 318 701058416H1 SOYMON033 g2606080 BLASTN 1385 1e−106 100 2352 318 701013289H1 SOYMON019 g2606080 BLASTN 1374 1e−105 99 2353 318 701002784H2 SOYMON019 g2606080 BLASTN 1365 1e−104 100 2354 318 700868516H1 SOYMON016 g2606080 BLASTN 1195 1e−103 100 2355 318 700978851H1 SOYMON009 g2606080 BLASTN 1325 1e−101 98 2356 318 701204954H1 SOYMON035 g2606080 BLASTN 770 1e−100 100 2357 318 700889102H1 SOYMON024 g2606080 BLASTN 1048 1e−100 99 2358 318 701053120H1 SOYMON032 g218332 BLASTN 1109 1e−100 90 2359 318 700731734H1 SOYMON010 g2606080 BLASTN 1308 1e−100 97 2360 318 700972625H1 SOYMON005 g2606080 BLASTN 1120 1e−98 99 2361 318 701006566H1 SOYMON019 g2606080 BLASTN 983 1e−97 99 2362 318 700952789H1 SOYMON022 g2606080 BLASTN 1276 1e−97 97 2363 318 701141518H1 SOYMON038 g2606080 BLASTN 716 1e−96 99 2364 318 700653475H1 SOYMON003 g3169543 BLASTN 1262 1e−96 87 2365 318 700650832H1 SOYMON003 g2606080 BLASTN 643 1e−95 97 2366 318 700678981H1 SOYMON007 g2606080 BLASTN 1142 1e−95 96 2367 318 700890311H1 SOYMON024 g2606080 BLASTN 1200 1e−95 100 2368 318 700892212H1 SOYMON024 g2606080 BLASTN 1250 1e−95 97 2369 318 700943424H1 SOYMON024 g2606080 BLASTN 1251 1e−95 99 2370 318 700833982H1 SOYMON019 g2606080 BLASTN 1255 1e−95 100 2371 318 700834361H1 SOYMON019 g2606080 BLASTN 981 1e−94 99 2372 318 700746379H1 SOYMON013 g2606080 BLASTN 1108 1e−94 96 2373 318 700889648H1 SOYMON024 g2606080 BLASTN 1238 1e−94 99 2374 318 701054868H1 SOYMON032 g2606080 BLASTN 1243 1e−94 95 2375 318 700959914H1 SOYMON022 g2606080 BLASTN 1226 1e−93 96 2376 318 701011518H1 SOYMON019 g2606080 BLASTN 705 1e−92 99 2377 318 700734053H1 SOYMON010 g2606080 BLASTN 765 1e−92 100 2378 318 701005295H1 SOYMON019 g2606080 BLASTN 962 1e−92 93 2379 318 700945690H1 SOYMON024 g2606080 BLASTN 1054 1e−92 99 2380 318 701118196H1 SOYMON037 g2606080 BLASTN 1100 1e−92 95 2381 318 700673512H1 SOYMON007 g2606080 BLASTN 1211 1e−92 97 2382 318 700852712H1 SOYMON023 g2606080 BLASTN 1215 1e−92 98 2383 318 701004755H1 SOYMON019 g2606080 BLASTN 1221 1e−92 99 2384 318 700677915H1 SOYMON007 g2606080 BLASTN 685 1e−91 99 2385 318 700977846H1 SOYMON009 g2606080 BLASTN 731 1e−91 99 2386 318 700831789H1 SOYMON019 g2606080 BLASTN 1204 1e−91 97 2387 318 700754901H1 SOYMON014 g2606080 BLASTN 1205 1e−91 100 2388 318 700666594H1 SOYMON005 g2606080 BLASTN 1210 1e−91 100 2389 318 700750890H1 SOYMON014 g2606080 BLASTN 1188 1e−90 99 2390 318 700890229H1 SOYMON024 g2606080 BLASTN 1195 1e−90 100 2391 318 700732660H1 SOYMON010 g2606080 BLASTN 1154 1e−89 95 2392 318 700764730H1 SOYMON023 g2606080 BLASTN 1181 1e−89 99 2393 318 701050015H1 SOYMON032 g218332 BLASTN 1185 1e−89 89 2394 318 700870180H1 SOYMON016 g2606080 BLASTN 710 1e−88 100 2395 318 701204236H2 SOYMON035 g2606080 BLASTN 904 1e−88 98 2396 318 700645782H1 SOYMON010 g2606080 BLASTN 633 1e−87 95 2397 318 700831711H1 SOYMON019 g2606080 BLASTN 1025 1e−87 96 2398 318 701056026H1 SOYMON032 g2606080 BLASTN 1158 1e−87 96 2399 318 700678853H1 SOYMON007 g2606080 BLASTN 1161 1e−87 97 2400 318 700852424H1 SOYMON023 g2606080 BLASTN 913 1e−86 95 2401 318 701049116H1 SOYMON032 g3169543 BLASTN 1146 1e−86 89 2402 318 700977788H1 SOYMON009 g2606080 BLASTN 642 1e−85 94 2403 318 700833546H1 SOYMON019 g2606080 BLASTN 1134 1e−85 94 2404 318 701004915H1 SOYMON019 g2606080 BLASTN 591 1e−84 96 2405 318 700730093H1 SOYMON009 g2606080 BLASTN 755 1e−84 96 2406 318 701119060H1 SOYMON037 g2606080 BLASTN 824 1e−84 97 2407 318 700963024H1 SOYMON022 g2606080 BLASTN 1116 1e−84 90 2408 318 700563532H1 SOYMON002 g22037 BLASTN 1116 1e−84 87 2409 318 700755891H1 SOYMON014 g2606080 BLASTN 1117 1e−84 94 2410 318 700850605H1 SOYMON023 g2606080 BLASTN 1118 1e−84 94 2411 318 700888245H1 SOYMON024 g2606080 BLASTN 643 1e−83 98 2412 318 701037091H1 SOYMON029 g2606080 BLASTN 821 1e−83 95 2413 318 700673790H1 SOYMON007 g2606080 BLASTN 1104 1e−83 95 2414 318 700845518H1 SOYMON021 g2606080 BLASTN 673 1e−82 91 2415 318 700854591H1 SOYMON023 g2606080 BLASTN 606 1e−81 95 2416 318 700907167H1 SOYMON022 g2606080 BLASTN 920 1e−81 96 2417 318 700978575H1 SOYMON009 g218332 BLASTN 971 1e−81 91 2418 318 700853484H1 SOYMON023 g2606080 BLASTN 1079 1e−81 92 2419 318 701124012H1 SOYMON037 g218332 BLASTN 1083 1e−81 89 2420 318 700835387H1 SOYMON019 g2606080 BLASTN 1087 1e−81 96 2421 318 700749133H1 SOYMON013 g2606080 BLASTN 571 1e−80 98 2422 318 700727185H1 SOYMON009 g2606080 BLASTN 730 1e−80 98 2423 318 700869024H1 SOYMON016 g2606080 BLASTN 807 1e−79 96 2424 318 701013537H1 SOYMON019 g2606080 BLASTN 929 1e−79 87 2425 318 701010402H1 SOYMON019 g218332 BLASTN 1055 1e−79 85 2426 318 701107955H1 SOYMON036 g2606080 BLASTN 1058 1e−79 87 2427 318 700731653H1 SOYMON010 g2606080 BLASTN 578 1e−78 94 2428 318 700888950H1 SOYMON024 g218332 BLASTN 765 1e−78 88 2429 318 700894112H1 SOYMON024 g2606080 BLASTN 842 1e−78 98 2430 318 701005565H1 SOYMON019 g2606080 BLASTN 1024 1e−78 92 2431 318 700548286H1 SOYMON002 g2606080 BLASTN 1045 1e−78 88 2432 318 700975854H1 SOYMON009 g22037 BLASTN 1053 1e−78 86 2433 318 700944525H1 SOYMON024 g218332 BLASTN 1054 1e−78 89 2434 318 701061312H1 SOYMON033 g2606080 BLASTN 773 1e−77 87 2435 318 700831277H1 SOYMON019 g2606080 BLASTN 947 1e−77 97 2436 318 700788482H1 SOYMON011 g2606080 BLASTN 1038 1e−77 89 2437 318 701055686H1 SOYMON032 g2606080 BLASTN 1039 1e−77 90 2438 318 701054768H1 SOYMON032 g2606080 BLASTN 786 1e−76 88 2439 318 700854891H1 SOYMON023 g2606080 BLASTN 1030 1e−76 93 2440 318 701215276H1 SOYMON035 g2606080 BLASTN 1030 1e−76 90 2441 318 700944860H1 SOYMON024 g2606080 BLASTN 887 1e−75 96 2442 318 701010957H1 SOYMON019 g2606080 BLASTN 1011 1e−75 87 2443 318 701007175H1 SOYMON019 g2606080 BLASTN 1013 1e−75 90 2444 318 700725567H1 SOYMON009 g2606080 BLASTN 1013 1e−75 93 2445 318 700904972H1 SOYMON022 g22037 BLASTN 1015 1e−75 89 2446 318 700747391H1 SOYMON013 g2606080 BLASTN 1017 1e−75 87 2447 318 700747523H1 SOYMON013 g22037 BLASTN 836 1e−74 86 2448 318 700561819H1 SOYMON002 g218332 BLASTN 999 1e−74 82 2449 318 700835961H1 SOYMON019 g218332 BLASTN 1006 1e−74 87 2450 318 700562318H1 SOYMON002 g2606080 BLASTN 986 1e−73 84 2451 318 700745092H1 SOYMON013 g2606080 BLASTN 987 1e−73 88 2452 318 700832618H1 SOYMON019 g2606080 BLASTN 975 1e−72 87 2453 318 700891092H1 SOYMON024 g2606080 BLASTN 982 1e−72 88 2454 318 701119264H1 SOYMON037 g2606080 BLASTN 690 1e−71 89 2455 318 700894436H1 SOYMON024 g2606080 BLASTN 901 1e−71 91 2456 318 700894532H1 SOYMON024 g22037 BLASTN 959 1e−71 89 2457 318 700891712H1 SOYMON024 g22037 BLASTN 960 1e−71 89 2458 318 700895985H1 SOYMON027 g2606080 BLASTN 964 1e−71 89 2459 318 701203243H1 SOYMON035 g2606080 BLASTN 969 1e−71 88 2460 318 700985945H1 SOYMON009 g218332 BLASTN 713 1e−70 90 2461 318 700984768H1 SOYMON009 g2606080 BLASTN 781 1e−69 84 2462 318 700675710H1 SOYMON007 g2606080 BLASTN 784 1e−69 91 2463 318 700829561H1 SOYMON019 g218332 BLASTN 935 1e−69 87 2464 318 700964918H1 SOYMON022 g22037 BLASTN 942 1e−69 83 2465 318 701046747H1 SOYMON032 g2606080 BLASTN 422 1e−68 84 2466 318 700745512H1 SOYMON013 g3169543 BLASTN 457 1e−68 85 2467 318 700666671H1 SOYMON005 g218332 BLASTN 506 1e−68 87 2468 318 700889555H1 SOYMON024 g3169543 BLASTN 930 1e−68 86 2469 318 701147844H1 SOYMON031 g3169543 BLASTN 932 1e−68 86 2470 318 701206247H1 SOYMON035 g3169543 BLASTN 934 1e−68 82 2471 318 701103801H1 SOYMON036 g218332 BLASTN 723 1e−67 88 2472 318 700943746H1 SOYMON024 g218332 BLASTN 913 1e−67 86 2473 318 700745956H1 SOYMON013 g22037 BLASTN 921 1e−67 83 2474 318 700893512H1 SOYMON024 g218332 BLASTN 835 1e−66 90 2475 318 700897675H1 SOYMON027 g22037 BLASTN 899 1e−66 83 2476 318 700565777H1 SOYMON002 g2606080 BLASTN 510 1e−65 89 2477 318 700749851H1 SOYMON013 g2606080 BLASTN 887 1e−65 89 2478 318 700746286H1 SOYMON013 g2606080 BLASTN 876 1e−64 82 2479 318 700869142H1 SOYMON016 g2606080 BLASTN 885 1e−64 100 2480 318 700892442H1 SOYMON024 g2606080 BLASTN 872 1e−63 84 2481 318 700964153H1 SOYMON022 g22037 BLASTN 873 1e−63 83 2482 318 700898176H1 SOYMON027 g3169543 BLASTN 873 1e−63 84 2483 318 701056245H1 SOYMON032 g218332 BLASTN 543 1e−61 84 2484 318 700835360H1 SOYMON019 g218332 BLASTN 839 1e−61 88 2485 318 700749067H1 SOYMON013 g3169543 BLASTN 473 1e−60 86 2486 318 701008962H1 SOYMON019 g3169543 BLASTN 614 1e−60 90 2487 318 700980315H1 SOYMON009 g3169543 BLASTN 655 1e−60 84 2488 318 701202680H1 SOYMON035 g2606080 BLASTN 678 1e−60 89 2489 318 701202364H1 SOYMON035 g2606080 BLASTN 711 1e−60 85 2490 318 701037195H1 SOYMON029 g218332 BLASTN 439 1e−59 86 2491 318 701011681H1 SOYMON019 g3169543 BLASTN 459 1e−59 83 2492 318 700976368H1 SOYMON009 g218332 BLASTN 363 1e−58 85 2493 318 700829847H1 SOYMON019 g218332 BLASTN 384 1e−58 86 2494 318 700561920H1 SOYMON002 g2606080 BLASTN 809 1e−58 88 2495 318 701004573H1 SOYMON019 g2606080 BLASTN 813 1e−58 77 2496 318 701049462H1 SOYMON032 g3169543 BLASTN 450 1e−57 82 2497 318 700866272H1 SOYMON016 g3169543 BLASTN 421 1e−54 77 2498 318 700892632H1 SOYMON024 g2606080 BLASTN 453 1e−54 84 2499 318 701215184H1 SOYMON035 g218332 BLASTN 464 1e−54 88 2500 318 700831177H1 SOYMON019 g2606080 BLASTN 759 1e−54 85 2501 318 700835115H1 SOYMON019 g2606080 BLASTN 762 1e−54 81 2502 318 701015056H1 SOYMON019 g3169543 BLASTN 447 1e−53 81 2503 318 700675496H1 SOYMON007 g2606080 BLASTN 465 1e−53 95 2504 318 701052767H1 SOYMON032 g2606080 BLASTN 753 1e−53 88 2505 318 700833078H1 SOYMON019 g3169543 BLASTN 414 1e−52 84 2506 318 700869165H1 SOYMON016 g3169543 BLASTN 534 1e−51 84 2507 318 700831532H1 SOYMON019 g2606080 BLASTN 655 1e−51 100 2508 318 701010104H2 SOYMON019 g2606080 BLASTN 698 1e−51 85 2509 318 700890513H1 SOYMON024 g22037 BLASTN 575 1e−50 88 2510 318 700890952H1 SOYMON024 g2606080 BLASTN 709 1e−50 75 2511 318 700567301H1 SOYMON002 g22037 BLASTN 716 1e−50 82 2512 318 700945284H1 SOYMON024 g3169543 BLASTN 701 1e−49 75 2513 318 701206626H1 SOYMON035 g3169543 BLASTN 702 1e−49 81 2514 318 700748456H1 SOYMON013 g2606080 BLASTN 384 1e−48 77 2515 318 700981883H1 SOYMON009 g2606080 BLASTN 419 1e−48 85 2516 318 700942575H1 SOYMON024 g22037 BLASTN 340 1e−46 82 2517 318 700945125H1 SOYMON024 g2606080 BLASTN 405 1e−46 81 2518 318 700830469H1 SOYMON019 g3169543 BLASTN 636 1e−44 83 2519 318 700991669H1 SOYMON011 g218332 BLASTN 630 1e−43 83 2520 318 700866064H1 SOYMON016 g3169543 BLASTN 453 1e−41 84 2521 318 700866806H1 SOYMON016 g218332 BLASTN 607 1e−41 96 2522 318 700893154H1 SOYMON024 g2606080 BLASTN 539 1e−38 87 2523 318 700893118H1 SOYMON024 g2606080 BLASTN 539 1e−38 87 2524 318 701142963H2 SOYMON038 g218332 BLASTN 569 1e−38 90 2525 318 700945968H1 SOYMON024 g218332 BLASTN 572 1e−38 86 2526 318 700945788H1 SOYMON024 g2606080 BLASTN 514 1e−36 90 2527 318 700563455H1 SOYMON002 g2606080 BLASTN 496 1e−32 83 2528 318 700888936H1 SOYMON024 g3169543 BLASTN 498 1e−32 86 2529 318 701039594H1 SOYMON029 g22037 BLASTN 254 1e−28 84 2530 318 701015024H1 SOYMON019 g218333 BLASTX 65 1e−14 66 2531 318 700893166H1 SOYMON024 g22037 BLASTN 232 1e−8 85 2532 4258 700646449H1 SOYMON013 g22037 BLASTN 584 1e−39 70 2533 4258 700952838H1 SOYMON022 g20373 BLASTN 557 1e−37 70 2534 4413 700902256H1 SOYMON027 g2606080 BLASTN 1215 1e−99 97 2535 4413 700900032H1 SOYMON027 g2606080 BLASTN 720 1e−95 98 2536 4413 701006182H1 SOYMON019 g2606080 BLASTN 1179 1e−89 99 2537 4413 700831710H1 SOYMON019 g2606080 BLASTN 1070 1e−80 97 2538 4413 701008850H1 SOYMON019 g2606080 BLASTN 999 1e−74 99 2539 4413 701015432H1 SOYMON019 g2606080 BLASTN 813 1e−68 95 2540 4413 700987094H1 SOYMON009 g2606080 BLASTN 928 1e−68 84 2541 4413 700736179H1 SOYMON010 g2606080 BLASTN 753 1e−63 96 2542 4413 700890230H1 SOYMON024 g2606080 BLASTN 798 1e−57 95 2543 4413 701015314H1 SOYMON019 g2606080 BLASTN 639 1e−49 97 2544 4413 701052019H1 SOYMON032 g2606080 BLASTN 448 1e−37 95 2545 4748 701209527H1 SOYMON035 g2606080 BLASTN 1207 1e−91 93 2546 4748 700561984H1 SOYMON002 g2606080 BLASTN 542 1e−81 94 2547 4748 700895166H1 SOYMON024 g2606080 BLASTN 1004 1e−74 98 2548 4748 700843735H1 SOYMON021 g2606080 BLASTN 227 1e−20 93 2549 869 700650545H1 SOYMON003 g2606080 BLASTN 804 1e−107 94 2550 869 701205255H1 SOYMON035 g2606080 BLASTN 1135 1e−101 98 2551 869 700562091H1 SOYMON002 g2606080 BLASTN 1311 1e−100 92 2552 869 701213906H1 SOYMON035 g2606080 BLASTN 1300 1e−99 100 2553 869 700567712H1 SOYMON002 g2606080 BLASTN 634 1e−95 97 2554 869 701010943H1 SOYMON019 g2606080 BLASTN 1236 1e−94 99 2555 869 701006976H1 SOYMON019 g2606080 BLASTN 601 1e−93 98 2556 869 700752409H1 SOYMON014 g2606080 BLASTN 1080 1e−92 100 2557 869 701204769H1 SOYMON035 g2606080 BLASTN 795 1e−90 100 2558 869 701042737H1 SOYMON029 g2606080 BLASTN 1058 1e−90 99 2559 869 700832091H1 SOYMON019 g2606080 BLASTN 1116 1e−88 99 2560 869 701049161H1 SOYMON032 g2606080 BLASTN 1053 1e−86 96 2561 869 700906541H1 SOYMON022 g2606080 BLASTN 1087 1e−86 96 2562 869 701008182H1 SOYMON019 g2606080 BLASTN 1111 1e−86 92 2563 869 700831609H1 SOYMON019 g2606080 BLASTN 611 1e−84 92 2564 869 700834954H1 SOYMON019 g2606080 BLASTN 835 1e−84 100 2565 869 701037284H1 SOYMON029 g2606080 BLASTN 858 1e−83 94 2566 869 700561458H1 SOYMON002 g2606080 BLASTN 1019 1e−83 93 2567 869 701208357H1 SOYMON035 g2606080 BLASTN 1113 1e−83 99 2568 869 700747138H1 SOYMON013 g2606080 BLASTN 985 1e−80 93 2569 869 701014835H1 SOYMON019 g2606080 BLASTN 891 1e−78 89 2570 869 700956359H1 SOYMON022 g2606080 BLASTN 1052 1e−78 96 2571 869 701012740H1 SOYMON019 g2606080 BLASTN 643 1e−77 93 2572 869 701042523H1 SOYMON029 g2606080 BLASTN 667 1e−74 95 2573 869 701205775H1 SOYMON035 g2606080 BLASTN 745 1e−74 100 2574 869 701049184H1 SOYMON032 g2606080 BLASTN 600 1e−72 95 2575 869 700889179H1 SOYMON024 g2606080 BLASTN 942 1e−69 92 2576 869 700963920H1 SOYMON022 g2606080 BLASTN 718 1e−66 90 2577 869 700737476H1 SOYMON010 g2606080 BLASTN 548 1e−44 97 2578 869 701044544H1 SOYMON032 g2606080 BLASTN 462 1e−43 96 2579 869 700737636H1 SOYMON010 g2606080 BLASTN 426 1e−34 95 2580 9398 700837013H1 SOYMON020 g2570066 BLASTN 1025 1e−76 88 2581 9398 700891526H1 SOYMON024 g2570066 BLASTN 868 1e−63 87 2582 14740 LIB3051-038- LIB3051 g2570066 BLASTN 1331 1e−102 86 Q1-K1-E10 2583 31182 LIB3051-015- LIB3051 g2570066 BLASTN 1540 1e−119 88 Q1-E1-F1 2584 318 LIB3050-024- LIB3050 g2606080 BLASTN 1736 1e−135 95 Q1-K1-H5 2585 318 LIB3050-012- L1B3050 g2606080 BLASTN 1564 1e−125 98 Q1-E1-F10 2586 318 LIB3056-013- LIB3056 g3169543 BLASTN 1617 1e−125 86 Q1-N1-H11 2587 318 LIB3028-026- LIB3028 g3169543 BLASTN 1393 1e−107 84 Q1-B1-F6 2588 318 LIB3049-031- LIB3049 g3169543 BLASTN 1290 1e−98 90 Q1 -E1-B6 2589 33428 LIB3051-085- LIB3051 g2570066 BLASTN 679 1e−53 86 Q1-K1-D11 2590 869 LIB3056-014- LIB3056 g2606080 BLASTN 1503 1e−132 96 Q1-N1-G8 SOYBEAN HEXOKINASE 2591 −700560085 700560085H1 SOYMON001 g1899024 BLASTN 456 1e−27 67 2592 −700752579 700752579H1 SOYMON014 g836808 BLASTX 113 1e−8 54 2593 −700753182 700753182H1 SOYMON014 g619928 BLASTX 234 1e−25 63 2594 −700838622 700838622H1 SOYMON020 g619927 BLASTN 767 1e−55 78 2595 −700840271 700840271H1 SOYMON020 g619927 BLASTN 525 1e−34 67 2596 −700844132 700844132H1 SOYMON021 g619927 BLASTN 474 1e−51 77 2597 −700898308 700898308H1 SOYMON027 g619927 BLASTN 464 1e−29 72 2598 −700904279 700904279H1 SOYMON022 g881521 BLASTX 129 1e−10 67 2599 −700904320 700904320H1 SOYMON022 g1899024 BLASTN 612 1e−42 71 2600 −700946357 700946357H1 SOYMON024 g619928 BLASTX 112 1e−18 69 2601 −700998007 700998007H1 SOYMON018 g1899024 BLASTN 367 1e−20 71 2602 −701097096 701097096H1 SOYMON028 g619927 BLASTN 488 1e−30 73 2603 −701102877 701102877H1 SOYMON028 g619927 BLASTN 551 1e−37 70 2604 −701103285 701103285H1 SOYMON028 g619928 BLASTX 179 1e−17 77 2605 −701105838 701105838H1 SOYMON036 g619928 BLASTX 274 1e−30 63 2606 −701138291 701138291H1 SOYMON038 g619927 BLASTN 819 1e−59 79 2607 12404 701065794H1 SOYMON034 g3087888 BLASTX 84 1e−11 44 2608 12404 701131030H1 SOYMON038 g1899025 BLASTX 120 1e−9 45 2609 12693 700846513H1 SOYMON021 g619927 BLASTN 459 1e−28 70 2610 12693 700656744H1 SOYMON004 g619927 BLASTN 251 1e−10 57 2611 12917 700906858H1 SOYMON022 g3087888 BLASTX 183 1e−32 80 2612 12917 700830011H1 SOYMON019 g619927 BLASTN 495 1e−32 70 2613 12917 701068501H1 SOYMON034 g619927 BLASTN 475 1e−29 72 2614 12917 701153981H1 SOYMON031 g3087887 BLASTN 440 1e−26 69 2615 222 700663332H1 SOYMON005 g619927 BLASTN 724 1e−51 76 2616 222 701142003H1 SOYMON038 g881520 BLASTN 542 1e−39 72 2617 222 700657213H1 SOYMON004 g881520 BLASTN 524 1e−34 73 2618 222 700833679H1 SOYMON019 g1899024 BLASTN 453 1e−28 80 2619 222 700556060H1 SOYMON001 g619927 BLASTN 463 1e−28 82 2620 23610 700984359H1 SOYMON009 g1899024 BLASTN 611 1e−42 73 2621 23610 701003284H1 SOYMON019 g1899024 BLASTN 577 1e−39 75 2622 25188 700760643H1 SOYMON015 g619927 BLASTN 701 1e−49 73 2623 25188 701056127H1 SOYMON032 g1899024 BLASTN 649 1e−45 70 2624 27316 701054167H1 SOYMON032 g3087888 BLASTX 177 1e−17 47 2625 27316 701054157H1 SOYMON032 g3087888 BLASTX 177 1e−17 47 2626 488 700682650H2 SOYMON008 g687676 BLASTN 730 1e−52 77 2627 488 700849894H1 SOYMON021 g687676 BLASTN 582 1e−39 76 2628 −GM32703 LIB3051-008- LIB3051 g1899024 BLASTN 981 1e−76 77 Q1 -E1-C12 2629 −GM9523 LIB3049-003- LIB3049 g619928 BLASTX 203 1e−37 64 Q1-E1-A6 2630 12693 LIB3051-106- LIB3051 g619927 BLASTN 459 1e−38 71 Q1-K1-A9 2631 488 L1B3040-006- L1B3040 g687676 BLASTN 622 1e−41 76 Q1 -E1-A12 2632 488 LIB3053-008- LIB3053 g687676 BLASTN 597 1e−39 75 Q1-N1-C6 2633 488 LIB3055-008- LIB3055 g687676 BLASTN 559 1e−36 75 Q1-N1-F5 2634 488 LIB3053-010- LIB3053 g687676 BLASTN 514 1e−32 75 Q1-N1-D8 SOYBEAN FRUCTOKINASE 2635 −700834049 700834049H1 SOYMON019 g1915974 BLASTX 112 1e−10 97 2636 −700905716 700905716H1 SOYMON022 g1915973 BLASTN 774 1e−55 77 2637 −700978126 700978126H1 SOYMON009 g1915973 BLASTN 565 1e−38 77 2638 −700983171 700983171H1 SOYMON009 g1915974 BLASTX 96 1e−9 93 2639 −701069652 701069652H1 SOYMON034 g297014 BLASTN 447 1e−27 73 2640 −701118004 701118004H2 SOYMON037 g2102690 BLASTN 440 1e−26 73 2641 −701209270 701209270H1 SOYMON035 g1052972 BLASTN 648 1e−45 79 2642 1174 700832430H1 SOYMON019 g1915973 BLASTN 638 1e−44 81 2643 1174 701101576H1 SOYMON028 g1915973 BLASTN 592 1e−40 79 2644 1174 700754333H1 SOYMON014 g1915973 BLASTN 323 1e−37 80 2645 1174 701004323H1 SOYMON019 g297014 BLASTN 560 1e−37 80 2646 1174 700988192H1 SOYMON009 g1915973 BLASTN 508 1e−33 78 2647 1174 700646337H1 SOYMON013 g1915974 BLASTX 153 1e−30 79 2648 1174 701039647H1 SOYMON029 g1915973 BLASTN 275 1e−12 80 2649 16472 701155250H1 SOYMON031 g1915973 BLASTN 642 1e−50 78 2650 16472 700953304H1 SOYMON022 g1915973 BLASTN 690 1e−48 79 2651 16472 700725996H1 SOYMON009 g1915973 BLASTN 362 1e−28 73 2652 17936 700965277H1 SOYMON022 g2102690 BLASTN 375 1e−42 77 2653 17936 700746240H1 SOYMON013 g2102690 BLASTN 606 1e−41 74 2654 22120 701215393H1 SOYMON035 g2102691 BLASTX 133 1e−11 86 2655 22586 701009695H1 SOYMON019 g2102690 BLASTN 696 1e−49 76 2656 22586 700900731H1 SOYMON027 g2102690 BLASTN 422 1e−26 76 2657 23551 701053585H1 SOYMON032 g2102691 BLASTX 120 1e−9 92 2658 28587 701156878H1 SOYMON031 g2102690 BLASTN 448 1e−33 72 2659 3876 700942858H1 SOYMON024 g297014 BLASTN 705 1e−49 74 2660 3876 701063105H1 SOYMON033 g1052972 BLASTN 679 1e−47 73 2661 3876 700844831H1 SOYMON021 g1915973 BLASTN 466 1e−37 72 2662 5530 700733713H1 SOYMON010 g1915974 BLASTX 156 1e−26 81 2663 5530 701057239H1 SOYMON033 g1915974 BLASTX 176 1e−17 92 2664 5530 700985231H1 SOYMON009 g297014 BLASTN 222 1e−16 79 2665 5805 701010614H1 SOYMON019 g1915973 BLASTN 958 1e−71 80 2666 5805 701003106H1 SOYMON019 g1915973 BLASTN 679 1e−64 81 2667 5805 700748895H1 SOYMON013 g1915973 BLASTN 475 1e−55 83 2668 5805 700892801H1 SOYMON024 g1915973 BLASTN 639 1e−55 80 2669 5805 700891914H1 SOYMON024 g1915973 BLASTN 639 1e−55 81 2670 5805 700962529H1 SOYMON022 g1915973 BLASTN 622 1e−54 82 2671 5805 700869294H1 SOYMON016 g1915973 BLASTN 760 1e−54 80 2672 5805 700986530H1 SOYMON009 g1915973 BLASTN 761 1e−54 80 2673 5805 700661115H1 SOYMON005 g1915973 BLASTN 682 1e−48 78 2674 5805 701041987H1 SOYMON029 g2970l4 BLASTN 475 1e−45 83 2675 5805 701006803H1 SOYMON019 g1915973 BLASTN 607 1e−41 80 2676 28587 LIB3028-008- LIB3028 g2102690 BLASTN 900 1e−66 68 Q1-B1-H3 2677 5530 LIB3055-004- LIB3055 g297014 BLASTN 606 1e−39 76 Q1-N1-H3 2678 5805 LIB3065-006- LIB3065 g1915973 BLASTN 954 1e−81 79 Q1-N1-F11 SOYBEAN NDP-KINASE 2679 33331 701108520H1 SOYMON036 g758643 BLASTN 473 1e−31 75 2680 23595 LIB3050-018- LIB3050 g758643 BLASTN 295 1e−13 76 Q1-E1-C4 2681 33331 LIB3040-037- LIB3040 g758643 BLASTN 413 1e−47 79 Q1-E1-D6 SOYBEAN GLUCOSE-6-PHOSPHATE 1-DEHYDROGENASE 2682 −700869140 700869140H1 SOYMON016 g2829880 BLASTX 164 1e−15 44 2683 −701065174 701065174H1 SOYMON034 g603219 BLASTX 86 1e−9 76 2684 −701130434 701130434H1 SOYMON037 g1197385 BLASTX 189 1e−19 55 2685 −701149522 701149522H1 SOYMON031 g603219 BLASTX 99 1e−8 71 2686 26484 701003905H1 SOYMON019 g1197385 BLASTX 138 1e−15 81 2687 9136 701038169H1 SOYMON029 g603219 BLASTX 139 1e−21 73 2688 9136 700903571H1 SOYMON022 g603219 BLASTX 144 1e−20 81 2689 9136 701045122H1 SOYMON032 g603219 BLASTX 100 1e−13 79 SOYBEAN PHOSPHOGLUCOMUTASE 2690 −700554424 700554424H1 SOYMON001 g534982 BLASTX 133 1e−25 60 2691 −700556670 700556670H1 SOYMON001 g3294468 BLASTN 355 1e−43 74 2692 −700563871 700563871H1 SOYMON002 g2795876 BLASTX 101 1e−16 75 2693 −700565101 700565101H1 SOYMON002 g3294466 BLASTN 588 1e−40 68 2694 −700566749 700566749H1 SOYMON002 g1814400 BLASTN 475 1e−41 73 2695 −700681382 700681382H2 SOYMON008 g3294467 BLASTX 98 1e−11 48 2696 −700763827 700763827H1 SOYMON018 g3192042 BLASTX 257 1e−29 60 2697 −700865583 700865583H1 SOYMON016 g3192042 BLASTX 134 1e−17 57 2698 −700891379 700891379H1 SOYMON024 g534982 BLASTX 167 1e−15 53 2699 −700942816 700942816H1 SOYMON024 g3294466 BLASTN 636 1e−44 74 2700 −701004954 701004954H1 SOYMON019 g1814400 BLASTN 790 1e−56 78 2701 −701011364 701011364H1 SOYMON019 g534982 BLASTX 284 1e−32 67 2702 −701057063 701057063H2 SOYMON033 g1814401 BLASTX 121 1e−9 60 2703 −701119491 701119491H1 SOYMON037 g1814400 BLASTN 762 1e−54 76 2704 −701149254 701149254H1 SOYMON031 g534982 BLASTX 147 1e−19 52 2705 10032 700988921H1 SOYMON011 g1814400 BLASTN 908 1e−66 80 2706 10032 701136003H1 SOYMON038 g1814400 BLASTN 842 1e−61 78 2707 10032 700953253H1 SOYMON022 g1814400 BLASTN 808 1e−58 77 2708 10032 701103083H1 SOYMON028 g1814400 BLASTN 813 1e−58 78 2709 10131 701104852H1 SOYMON036 g3294466 BLASTN 302 1e−27 74 2710 10131 700970420H1 SOYMON005 g2829893 BLASTX 240 1e−26 56 2711 1180 701125681H1 SOYMON037 g2829893 BLASTX 163 1e−15 82 2712 1180 700559947H1 SOYMON001 g2829893 BLASTX 163 1e−15 82 2713 1180 700556009H1 SOYMON001 g2829893 BLASTX 102 1e−14 87 2714 13262 701006086H2 SOYMON019 g3294466 BLASTN 734 1e−52 75 2715 13262 701137937H1 SOYMON038 g3294466 BLASTN 491 1e−32 71 2716 13262 701004207H1 SOYMON019 g3294466 BLASTN 271 1e−30 75 2717 13262 700904551H1 SOYMON022 g3294466 BLASTN 473 1e−30 75 2718 13262 701014357H1 SOYMON019 g1814401 BLASTX 210 1e−21 83 2719 13262 701146638H1 SOYMON031 g1814401 BLASTX 111 1e−20 80 2720 13262 700833416H1 SOYMON019 g1814400 BLASTN 374 1e−20 74 2721 13262 701148967H1 SOYMON031 g1814401 BLASTX 194 1e−19 82 2722 13262 701156042H1 SOYMON031 g1814401 BLASTX 182 1e−18 67 2723 13262 700943365H1 SOYMON024 g1814401 BLASTX 168 1e−16 76 2724 13262 701105762H1 SOYMON036 g1814401 BLASTX 165 1e−15 83 2725 13262 701038338H1 SOYMON029 g1814400 BLASTN 186 1e−13 77 2726 13262 700645989H1 SOYMON011 g1814401 BLASTX 133 1e−11 78 2727 13262 700868941H1 SOYMON016 g1814400 BLASTN 181 1e−9 80 2728 19312 701121150H1 SOYMON037 g3294468 BLASTN 501 1e−61 79 2729 19312 700742959H1 SOYMON012 g3294468 BLASTN 440 1e−45 83 2730 19312 701135418H1 SOYMON038 g3294468 BLASTN 459 1e−42 79 2731 19312 700979514H2 SOYMON009 g1814400 BLASTN 612 1e−42 78 2732 19883 701133631H2 SOYMON038 g1814400 BLASTN 758 1e−54 75 2733 19883 700970758H1 SOYMON005 g1814400 BLASTN 717 1e−50 77 2734 19883 701153416H1 SOYMON031 g1814400 BLASTN 691 1e−48 76 2735 26278 701214005H1 SOYMON035 g534982 BLASTX 118 1e−8 47 2736 −GM1647 LIB3028-009- LIB3028 g534982 BLASTX 192 1e−42 57 Q1-B1-F3 2737 −GM17162 LIB3055-012- LIB3055 g1814400 BLASTN 491 1e−29 62 Q1-N1-B3 2738 13262 LIB3028-003- LIB3028 g1814400 BLASTN 1069 1e−80 76 Q1-B1-B11 2739 13262 LIB3054-009- LIB3054 g1814400 BLASTN 612 1e−40 73 Q1-N1-A12 2740 13262 LIB3054-009- LIB3054 g1814401 BLASTX 200 1e−36 73 Q1-N1-A5 SOYBEAN UDP-GLUCOSE PYROPHOSPHORYLASE 2741 −700665357 700665357H1 SOYMON005 g1388021 BLASTX 183 1e−18 69 2742 −700674325 700674325H1 SOYMON007 g218000 BLASTN 645 1e−44 72 2743 −700835903 700835903H1 SOYMON019 g1388021 BLASTX 135 1e−11 68 2744 −700841466 700841466H1 SOYMON020 g1388021 BLASTX 115 1e−14 56 2745 −700846570 700846570H1 SOYMON021 g3107930 BLASTN 486 1e−31 70 2746 −700888547 700888547H1 SOYMON024 g3107930 BLASTN 582 1e−39 81 2747 −700973436 700973436H1 SOYMON005 g1212996 BLASTX 132 1e−15 51 2748 −700985779 700985779H1 SOYMON009 g3107930 BLASTN 958 1e−71 83 2749 −700992994 700992994H1 SOYMON011 g1388021 BLASTX 103 1e−10 64 2750 −701061122 701061122H1 SOYMON033 g1388021 BLASTX 129 1e−19 73 2751 −701063465 701063465H1 SOYMON033 g3107930 BLASTN 426 1e−62 82 2752 −701118256 701118256H1 SOYMON037 g3107930 BLASTN 378 1e−31 83 2753 11810 700952705H1 SOYMON022 g3107930 BLASTN 613 1e−54 80 2754 11810 701060568H1 SOYMON033 g3107930 BLASTN 652 1e−45 81 2755 11810 701002783H2 SOYMON019 g3107930 BLASTN 458 1e−43 80 2756 11810 701202674H1 SOYMON035 g218000 BLASTN 331 1e−33 73 2757 11810 700871590H1 SOYMON018 g218000 BLASTN 326 1e−27 76 2758 11810 700747279H1 SOYMON013 g1388021 BLASTX 154 1e−21 75 2759 11810 701014424H1 SOYMON019 g1388021 BLASTX 131 1e−20 84 2760 11810 701039454H1 SOYMON029 g1388021 BLASTX 157 1e−16 80 2761 11810 701054271H1 SOYMON032 g1388021 BLASTX 154 1e−15 71 2762 11810 700955092H1 SOYMON022 g1388021 BLASTX 154 1e−14 71 2763 11810 701107189H1 SOYMON036 g1388021 BLASTX 155 1e−14 72 2764 11810 701107930H1 SOYMON036 g218000 BLASTN 308 1e−14 75 2765 11810 700904384H1 SOYMON022 g1388021 BLASTX 149 1e−13 72 2766 11810 700729516H1 SOYMON009 g1388021 BLASTX 155 1e−13 72 2767 11810 701009325H1 SOYMON019 g1388021 BLASTX 143 1e−12 75 2768 11821 701060627H1 SOYMON033 g218000 BLASTN 253 1e−26 74 2769 11821 701004671H1 SOYMON019 g21599 BLASTX 166 1e−22 77 2770 11821 700964889H1 SOYMON022 g1388021 BLASTX 167 1e−16 67 2771 13178 700562308H1 SOYMON002 g3107930 BLASTN 1198 1e−91 87 2772 13178 701049018H1 SOYMON032 g3107930 BLASTN 1101 1e−82 88 2773 13178 701126215H1 SOYMON037 g3107930 BLASTN 1072 1e−80 88 2774 13178 701211745H1 SOYMON035 g3107930 BLASTN 1038 1e−77 87 2775 13178 700850417H1 SOYMON023 g3107930 BLASTN 1022 1e−76 87 2776 13178 700665292H1 SOYMON005 g3107930 BLASTN 980 1e−72 88 2777 13178 700994009H1 SOYMON011 g3107930 BLASTN 958 1e−71 86 2778 13178 700895203H1 SOYMON024 g3107930 BLASTN 864 1e−68 86 2779 13178 701151725H1 SOYMON031 g3107930 BLASTN 800 1e−66 87 2780 13178 700988803H1 SOYMON011 g3107930 BLASTN 896 1e−65 80 2781 13178 700646581H1 SOYMON014 g3107930 BLASTN 483 1e−62 81 2782 13178 701153726H1 SOYMON031 g3107930 BLASTN 832 1e−60 87 2783 13178 701152333H1 SOYMON031 g3107930 BLASTN 674 1e−56 79 2784 13178 700756960H1 SOYMON015 g3107930 BLASTN 787 1e−56 86 2785 13178 700556901H1 SOYMON001 g218000 BLASTN 772 1e−55 84 2786 13178 701063605H1 SOYMON033 g3107930 BLASTN 566 1e−51 89 2787 13178 701212385H1 SOYMON035 g3107930 BLASTN 390 1e−23 78 2788 13178 700889518H1 SOYMON024 g3107931 BLASIX 131 1e−10 64 2789 17057 700740176H1 SOYMON012 g3107930 BLASTN 798 1e−57 81 2790 17057 700905747H1 SOYMON022 g3107930 BLASTN 511 1e−33 81 2791 1955 701059208H1 SOYMON033 g3107930 BLASTN 1034 1e−77 85 2792 1955 700984109H1 SOYMON009 g3107930 BLASTN 970 1e−72 84 2793 1955 701209482H1 SOYMON035 g3107930 BLASTN 931 1e−68 84 2794 1955 700554847H1 SOYMON001 g3107930 BLASTN 493 1e−66 83 2795 1955 701150363H1 SOYMON031 g3107930 BLASTN 898 1e−66 84 2796 1955 700986014H1 SOYMON009 g3107930 BLASTN 907 1e−66 84 2797 1955 700564270H1 SOYMON002 g3107930 BLASTN 501 1e−65 84 2798 1955 700844253H1 SOYMON021 g3107930 BLASTN 875 1e−64 84 2799 1955 701140892H1 SOYMON038 g3107930 BLASTN 879 1e−64 83 2800 1955 700685893H1 SOYMON008 g3107930 BLASTN 832 1e−60 81 2801 1955 700789732H1 SOYMON011 g3107930 BLASTN 554 1e−57 84 2802 1955 700902418H1 SOYMON027 g3107930 BLASTN 731 1e−52 83 2803 1955 701128306H1 SOYMON037 g3107930 BLASTN 466 1e−46 82 2804 1955 701057973H1 SOYMON033 g3107930 BLASTN 422 1e−43 78 2805 21035 700946288H1 SOYMON024 g3107931 BLASTX 175 1e−17 72 2806 21035 701043539H1 SOYMON029 g3107931 BLASTX 147 1e−13 71 2807 30564 701063642H1 SOYMON033 g3107931 BLASTX 181 1e−25 75 2808 −GM18453 LIB3065-001- LIB3065 g1212996 BLASTX 68 1e−29 57 Q1-N1-H4 2809 −GM32502 LIB3051-013- LIB3051 g3107931 BLASTX 227 1e−47 51 Q1-E1-A6 2810 11810 LIB3030-010- LIB3030 g21598 BLASTN 1115 1e−84 76 Q1-B1-H12 2811 13178 LIB3056-014- LIB3056 g3107930 BLASTN 1145 1e−99 84 Q1-N1-G7 2812 1955 LIB3056-012- LIB3056 g3107930 BLASTN 856 1e−62 80 Q1-N1-D4 2813 30564 LIB3050-003- LIB3050 g3107930 BLASTN 1078 1e−81 74 Q1-E1-D8 2814 30564 LIB3050-010- LIB3050 g3107930 BLASTN 1050 1e−78 75 Q1-E1-D6 *Table Headings Cluster ID

A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. The cluster-ID entries in the table refer to the cluster with which the particular clone in each row is associated.

Clone ID

The clone ID number refers to the particular clone in the PhytoSeq database. Each clone ID entry in the table refers to the clone whose sequence is used for (1) the sequence comparison whose scores are presented and/or (2) assignment to the particular cluster which is presented. Note that a clone may be included in this table even if its sequence comparison scores fail to meet the minimum standards for similarity. In such a case, the clone is included due solely to its association with a particular cluster for which sequences of one or more other member clones possess the required level of similarity.

Library

The library ID refers to the particular cDNA library from which a given clone is obtained. Each cDNA library is associated with the particular tissue(s), line(s) and developmental stage(s) from which it is isolated.

NCBI gi

Each sequence in the GenBank public database is arbitrarily assigned a unique NCBI gi (National Center for Biotechnology Information GenBank Identifier) number. In this table, the NCBI gi number which is associated (in the same row) with a given clone refers to the particular GenBank sequence which is used in the sequence comparison. This entry is omitted when a clone is included solely due to its association with a particular cluster.

Method

The entry in the “Method” column of the table refers to the type of BLAST search that is used for the sequence comparison. “CLUSTER” is entered when the sequence comparison scores for a given clone fail to meet the minimum values required for significant similarity. In such cases, the clone is listed in the table solely as a result of its association with a given cluster for which sequences of one or more other member clones possess the required level of similarity.

Score

Each entry in the “Score” column of the table refers to the BLAST score that is generated by sequence comparison of the designated clone with the designated GenBank sequence using the designated BLAST method. This entry is omitted when a clone is included solely due to its association with a particular cluster. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

P-Value

The entries in the P-Value column refer to the probability that such matches occur by chance.

% Ident

The entries in the “% Ident” column of the table refer to the percentage of identically matched nucleotides (or residues) that exist along the length of that portion of the sequences which is aligned by the BLAST comparison to generate the statistical scores presented. This entry is omitted when a clone is included solely due to its association with a particular cluster. 

1. A substantially purified nucleic acid molecule that encodes a maize or a soybean enzyme, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs:11, 446, 935, 1108, 2042, 2166, 2252, 2644, 2681, and 2753, wherein said enzyme encoded by said nucleic acid molecule is triose phosphate isomerase, vacuolar H⁺ translocating-pyrophosphatase, sucrose synthase, hexokinase, fructose 1,6-bisphosphate aldolase, fructose 6-phosphate 2-kinase, invertase, fructokinase, NDP-kinase, and UDP-glucose pyrophosphorylase, respectively.
 2. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a maize triose phosphate isomerase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 11. 3. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a maize vacuolar H⁺ translocating-pyrophosphatase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 446. 4. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a maize sucrose synthase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 935. 5. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a maize hexokinase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 1108. 6. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy fructose 1,6-bisphosphate aldolase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2042. 7. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy fructose 6-phosphate 2-kinase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2166. 8. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy invertase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2252. 9. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy fructokinase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2644. 10. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy NDP-kinase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2681. 11. The substantially purified nucleic acid molecule of claim 1, wherein said enzyme is a soy UDP-glucose pyrophosphorylase, and wherein said nucleic acid sequence shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2753. 12. A substantially purified nucleic acid molecule comprising a nucleic acid sequence which shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11, 446, 935, 1108, 2042, 2166, 2252, 2644, 2681, and
 2753. 13. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares bewteen 100% and 95% sequence identity with a nucleic acid sequence of SEQ ID NO:
 11. 14. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 446. 15. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 935. 16. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 1108. 17. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2042. 18. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2166. 19. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2252. 20. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2644. 21. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid sequence which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2681. 22. The substantially purified nucleic acid molecule of claim 12, wherein said nucleic acid molecule comprises a nucleic acid which shares between 100% and 95% sequence identity with the nucleic acid sequence of SEQ ID NO:
 2753. 23. A transformed seed comprising a transformed plant cell comprising a nucleic acid molecule which comprises (a) an exogenous promoter region which functions in said plant cell to cause the production of an mRNA molecule, which is linked to; (b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11,446,935, 1108, 2042, 2166, 2252, 2644, 2681, 2753 and complements thereof, which is linked to; (c) a 3’ non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3’ end of said mRNA molecule.
 24. The transformed seed according to claim 23, wherein said nucleic acid sequence shares 100% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11, 446, 935, 1108, 2042, 2166, 2252, 2644, 2681, 2753 and complements thereof.
 25. A method of growing a transgenic plant comprising (a) planting a transformed seed comprising a nucleic acid molecule, which comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11, 446, 935, 1108, 2042, 2166, 2252, 2644, 2681, 2753, and complements thereof; and (b) growing a plant from said seed. 